Preparation and methods of use for ortho-aryl 5-membered heteroaryl-carboxamide containing multi-targeted kinase inhibitors

ABSTRACT

The present disclosure relates to compounds of the Formula (I): 
                         
and pharmaceutically acceptable salts, as kinase modulators, compatible with the Type-II inhibition of kinases.

RELATED APPLICATIONS

This application a Continuation of U.S. patent application Ser. No.14/173,125, filed Feb. 5, 2014, which is a Continuation-in-Part ofPCT/US2012/049559, filed Aug. 3, 2012, which claims the benefit of U.S.Provisional Application No. 61/515,434, filed Aug. 5, 2011. The entirecontents of each of these applications are expressly incorporated hereinby reference.

FIELD OF THE INVENTION

The present disclosure is directed to novel kinase inhibitors, processesfor their preparation, pharmaceutical compositions containing theseinhibitors, and their use in the treatment of various diseases andconditions.

BACKGROUND

Roles and Actions of Kinases:

Receptor protein kinases coupled with complex downstream kinase andphosphatase mediated cascades and feedback loops play critical roles insignal transduction from the cell exterior into the nucleus wheretranscriptional regulation takes place. Upon receptor activation, thissignal transduction is performed by the act of phosphate transfer to theserine, threonine, and tyrosine residues of proteins that results inenzyme activation or inactivation, changes in conformation, increased ordecreased affinity for other proteins, appropriate localization, and insome cases targeting of proteins for degradation by the proteosome.Understandably, these processes are tightly regulated and balancedthrough control of external receptor ligands as well as expression ofreceptors, receptor antagonists, decoy receptors, and throughredundancies or crosstalk between signaling pathways. Thus, it isaccepted that kinases perform essential functions in regulating cellgrowth and proliferation, differentiation and cell development, celldivision and aberrant mitogenesis. Kinases also mediate and regulatecell adhesion, angiogenesis, stress responses, cell-cell or cell-matrixinteractions, and short range contact-mediated axional guidance.Mechanistically similar non-protein kinases, such as PI3Ks and SPK1, canalso phosphorylate other ligands that contribute to the regulatoryprocess (Brown J. R., BMC Evolutionary Biology (2011) 11(4): 1471-2148;Alvarez S. E., Nature (2010) 465: 1084-1088). Therefore, diseases andconditions where aberrant kinase activity plays a role are plentiful.However, the complexities of the systems biology combined with thestructural homology of the kinase sites in the over 500 members of thehuman kinome presents a significant challenge for disease specificintervention by kinase inhibitors.

The Therapeutic Utility of Kinase Inhibitors:

With the advent of Imatinib (Deininger M., Blood (2005) 105(7):2640-2653) the primary focus for kinase inhibitor development hasbeen for the targeted treatment of specific cancers where mutationdriven aberrant kinase activations are particularly significant.Applications for kinase inhibitors in cancer therapy continues to evolveand these utilities have been extensively reviewed (Zhang J., Nat. Rev.Cancer (2009) 9(1): 28-39). However, strong links exist between cancerprogression and a pro-growth inflammatory environment have beenestablished (Rakoff-Nahoum S., Yale J. Biol. Med. (2006), 79:123-130;Schmid M. C., Cancer Cell, (2011) 19(6): 715-727). In addition, adiverse set of kinases participate in chronic inflammatory diseases,such as rheumatoid arthritis, psoriatic arthritis, inflammatory boweldisease, and chronic obstructive pulmonary disease, which are highlydebilitating diseases that affect a large segment of our population.Moreover, it has become apparent that metabolic diseases such as type 2diabetes, neuro-degenerative disorders such as Alzheimers, andcardiovascular diseases such as athlerosclerosis, also have a stronginflammatory component involving overactive kinase pathways. Therefore,selective inhibition of key kinases and their compensatory mechanismscontinues to be pursued as a promising strategy for therapeuticintervention. (Garuti L., Current Medicinal Chemistry (2010) 17:2804-2821).

Due to the complexity of signal transduction pathways, compensatorymechanisms often confound the initial therapeutic benefits seen withhighly selective targeted kinase inhibitors. Conversely, undesiredoff-target effects can introduce significant toxicity. The ongoingchallenge in the development of kinase inhibitors, particularly forchronic administration, is achieving the balance between efficacy andsafety. Since, the aberrant activity of kinases is fundamental to manychronic diseases and cancers, much effort continues to be expended tounderstand their diverse and complex roles in basic physiology.

Kinases in Inflammatory Diseases:

Mitogen-activated protein (MAP) kinases are known to play key roles inthe transmission of signals from cell surface receptors to transcriptionfactors which up-regulate the expression of pro-inflammatory cytokines.The MAP kinase p38-α is participant in one pathway that regulates theproduction of the pro-inflammatory cytokines TNF-α, IL-1, IL-6 and IL-8,as well as the enzymes COX-2, MMP-1 and MMP-3. It has also beendemonstrated that inhibiting p38-α kinase delays the onset of jointdisease in animal models of arthritis (Mihara K., British Journal ofPharmacology (2008) 154:153-164) by arresting the over production ofthese pro-inflammatory cytokines (Schindler J. F. J. Dental Res. (2007),86(9): 800-811). However, high hopes for p38 inhibitors as a singletarget therapy for chronic inflammation have not been realized inclinical studies which demonstrated these effects to be short lived,presumably by activation of compensatory mechanisms (Sweeney S. E.,Nature Reviews Rheumatology (2009) 5: 475-477). However, recent reportsof phase-II data in osteoarthritic patients using a sustained releaseformulations of p38 inhibitor, FX005, delivered intra-articularly to theknee look promising for both relief of pain and inflammation. Morerecently, redundant and non-redundant functions of the JNK isoforms JNK1and JNK2 in the immune system and arthritis have been described (GumaM., Proc Natl Acad Sci USA. (2010), 107(51):22122-7; Hommes D.,Gastroenterology. (2002), 122(1):7-14. Stambe C., Kidney Int (2003),64:2121-2132; Ma F. Y., Laboratory Investigation (2009) 89: 470-484).Additionally, inhibitors of Janus family kinases (JAK1, JAK2, and JAK3)have demonstrated anti-inflammatory effects in animal models (Stump KL., Arthritis Research and Therapy (2011) 13:R68; Meyer D M., J.Inflammation (2010) 7(41):1-12). Consequently, interest insmall-molecule therapeutics that target p38, JAK, and JNK isoforms forinflammatory diseases remains high (Liu C., J. Med. Chem. (2010) 53(18):6629-6639).

Encouraging anti-inflammatory preclinical and clinical results withImatinib, the well-known anti-cancer kinase inhibitor with Abl, PDGFR,c-KIT, and c-Raf activities, (Deininger M., Blood (2005) 105(7):2640-2653.) has rekindled interest in the development of kinaseinhibitors as anti-inflammatory agents (Iyoda M., Kidney International(2009), 75(10):1060-70; Ghofrani H. A., J Am Coll Cardiol (2009)54:108-117; Louvet C, Proc. Natl. Acad. Sci. USA (2008)105:18895-18900). Both protein and lipid kinases are now seen aspotential targets for the attenuation of the inflammatory response.Macrophage colony stimulating factor receptor (CSF-1R or FMS) along withKIT, FLT3, and PDGFR-a/b, are members of the type-III receptor tyrosinekinase family which have enjoyed much attention as potential kinasetargets (Tamura T. and Koch A., Anti-Inflamm Anti-Allergy Agents in Med.Chem. (2007) 6: 47-60). CSF-1R and its ligand (CSF-1) have beenimplicated in a range of macrophage and osteoclast related pathologicalprocesses, including rheumatoid arthritis, osteo-arthritis, progressionof atherosclerotic plaques, and bone metastasis (Ohno H. et al., Mol.Cancer Ther. (2006) 5(11): 2634-2643). c-Kit, the receptor of stem cellfactor (SCF), plays a key role in modulation of histamine release frommast cells and influences cell migration and adhesion to theextracellular matrix Inhibition of c-Kit mediates signaling in cynovialtissue from patients with rheumatoid arthritis and induced apoptosis ofmast cells. The platelet derived growth factor (PDGF) receptor, which isstructurally related to both CSF-1R and KIT, is important for theproliferation and migration of mesenchymal cells and is thought to playa role in the airway remodeling in asthma patients, inflammation inarthritis, and psoriasis. Additionally, Raf-1(c-Raf) inhibition has beenshown to suppress smoke-induced airway hyperresponsiveness in mice (LieY. et al., Respiratory. Res. (2008) 9(71): 1-10) and has been associatedwith clinical remission is severe Crohn's disease (Lowenberg M. et al.,J. Immunol. (2005) 175:2293-2300).

Additionally, inhibition of the neurotrophin/Trk pathway using NGFantibodies or non-selective small molecule inhibitors of Trk A, B and Chas been reported to be effective in treatment of pre-clinical models ofinflammatory diseases such as asthma, interstitial cystitis,inflammatory bowel disease, atopic dermatitis and psoriasis(Freund-Michel V., Pharmacology & Therapeutics (2008), 117(1): 52-76; HuV., The Journal of Urology (2005), 173(3): 1016-21; Di MoIa F. F., Gut(2000), 46(5), 670-678; Dou Y-C., Archives of Dermatological Research(2006) 298(1):31-37. Raychaudhuri S. P., Investigative Dermatology(2004), 122(3): 812-819). PI3K-γ and PI3K-δ have been stronglyimplicated as a major player in inflammatory conditions (Ruckle T., Nat.Rev. Drug Disc. (2006) 5:903-918; Hawkins P. T., Science (2007)318:64-66; Barberis L., Thromb Haemost (2008) 99: 279-285.) and tumorgrowth in a model of colitis-associated cancer (Gonzalez-Garcia A.,Gastroenterology (2010) 138:1374-1383). The links between inflammationand proliferative diseases also points to the potential ofanti-inflammatory agents as an adjunct to cancer therapy (Karin M.,Proc. Am. Thor. Soc. (2005) 2: 368-390; Rakoff-Nahoum s., J. Biol. Med.(2006), 79:123-130; Gust T. C., Anti-Inflammatory & Anti-Allergy Agentsin Medicinal Chemistry, (2007), 6:19-27; Schmid M. C., Cancer Cell(2011) 19, 715-727). PI3Kγ-deficient mice phenotypes suggest a widevariety of potential therapeutic applications for a selective inhibitor,including: allergic hyper-responsiveness, anaphylaxis, thrombosis,rheumatoid arthritis, glomerulonephritis, systemic lupus erythematosus(SLE), lung injury and airway inflammation related conditions such asCOPD, pancreatitis, reduced contractility due to heart failure andischaemia, and hypertension. Due to system redundancies and the lack ofinvolvement of PI3Kγ in metabolism and house keeping functions, effectsinduced by PI3Kγ inhibition, which may involve inhibition of chemotaxisand cell specific functions, are seen as soft methods of interventionwhere undesirable side effects may be minimized (Ruckle T., Nat. Rev.Drug Disc. (2006) 5:903-918).

Neurodegenrative Diseases:

Several kinases are believed to play a role in the pathogenesis of manyneurodegenerative disorders. For example, the active form of c-Raf (alsotermed Raf-1) is upregulated in the brains of Alzheimer's patients andin transgenic Alzheimer's mouse models. The persistent activation ofcRaf-1 can activate NFκB and consequently, upregulate the expression ofseveral of its downstream factors such as the amyloid precursor protein(APP), Cox-2 and iNOS. These factors have been found upregulated innumerous neurodegenerative conditions including Alzheimer's, epilepsy,brain trauma, and psychological stress (Burgess S., CNS Neurol DisordDrug Targets. (2010) 1:120-7). In addition, the neurotrophin/Trkpathway, particularly through BDNF/TrkB signaling, has been linked tothe etiology of neurodegenerative diseases including Parkinson'sdisease, multiple sclerosis, and Alzheimer's Disease (AD) (Sohrabji F.,Frontiers in Neuroendocrinology (2006), 27(4), 404-414). Several kinasesthought to be involved in the underlying inflammatory cause of AD,including GSK3, DAPK1, MAP-kinase, MLCK, and ROCK-1, have been studied.(Villar-Cheda B., Neurobiol Dis. 2012 47(2):268-79) and recentlyinhibitors of cytokin production in the brain have shown success inmodels of this disease (Bachstetter A. D., J. Neuroscience, 25 Jul.2012, 32(30): 10201-10210). Parkinson's Disease (PD) has links to someof the se kinases but the over expression and mutations in LRRK2 hasstimulated efforts to inhibit this kinase as a primary target (Kramer T,ACS Chem. Neurosci., (2012), 3(3), 151-160).

Infectious Disease:

The TrkA receptor kinase has been reported to be critical to the diseaseprocess in the parasitic infection of Trypanosoma cruzi (Chagas disease)in human hosts (de Melo-Jorge M., Cell Host & Microbe (2007)1(4):251-261).). Furthermore, a recent study demonstrated in mice thatadministration of the broad-spectrum receptor tyrosine kinase inhibitorsunitinib blocked the vascular remodeling and progressive splenomegalyassociated with experimental visceral leishmaniasis (Dalton J. E., JClin Invest. (2010) 120(4):1204-1216). In addition, sunitinib treatmentrestored the integrity of the splenic microarchitecture. While thistreatment alone was insufficient to cause a reduction in tissue parasiteburden, sunitinib proved to be successful as an adjunct therapy byproviding dose-sparing effects when combined with an immune-dependentanti-leishmanial drug. These data suggest, therefore, thatmulti-targeted tyrosine kinase inhibitors may prove clinically useful asagents in the treatment of parasitic infections and perhaps otherinfectious diseases.

Diabetes:

Imatinib and other TKIs counteract diabetes not only in non-obesediabetic mice, but also in streptozotocin diabetic mice, db/db mice,high-fat treated rats and humans with Type-2 diabetes (T2D). In thelater stages of T2D, β-cells are damaged. The molecular events leadingto cytokine-induced β-cell dysfunction and death have been linked to theactivation of the transcription factors NF-κB (nuclear factor κB) andSTATs (signal transducers and activators of transcription). The MAPKs(mitogen-activated protein kinases), such as JNK (c-Jun N-terminalkinase) and p38 MAPK, in response to both cytokines and oxidativestress, play a central role in this chain of events (Eizirik M.,Diabetologia (2001) 44(12):2115-2133). Although the mechanisms ofprotection need to be investigated further, the effects of imatinib andother TKIs in human T2D and the rapidly growing findings from animalmodels of Type 1 diabetes (T1D) and T2D are encouraging and give hope toimproved treatment of this disease (Dariush, 2010). In addition to NF-κBand p38 pathway involvement, PI3K pathways have been implicated as asignaling pathway involved in LPS induced TNF-alpha production in humanadipocytes (Hoareau L., Journal of Inflammation (2010) 7:1-12).

Cardiovascular Disease:

Atherosclerosis has also been linked to DDR1 and DDR2 expression.Neointimal thickening is a major cause of restenosis and atherosclerosisand smooth muscle cells (SMCs) are the predominant cell typecontributing to its formation after arterial injury. Collagen synthesisby the SMCs after arterial injury acts as an important regulator of thearterial repair through DDR1 and DDR2 activation. In non-human primatehypercholesterolemic diet studies, both DDRs were found to be highlyexpressed by smooth muscle cells (SMCs) in the fibrous cap ofatherosclerotic plaques (Ferri N, Am J Pathol, 2004, 164:1575-1585).Shyu et al demonstrated in balloon injury rat carotid artery model thatDDR2 directly promoted the migration and proliferation of vascularsmooth muscle cells which contributed to the development of neointimalformation in restenosis and accelerated the arteriopathy. Their studydemonstrated that siRNA-mediated inhibition of DDR2 protein expressionat the time of balloon injury reduced the neointimal lesion areasignificantly (Shyu K G, Arterioscler Thromb Vase Biol. 2008;28:1447-1453). Furthermore, DDR1 null mice had much less severe collagenaccumulation in atherosclerotic plaques than wild type mice which wasattributed to their considerably lower expression of MMP2 and decreasedSMC proliferation and migration (Hou G, J Clin Invest. 2001, 107(6):727;Hou G, Circ. Res 2002, 90:1147-1149).

Several growth factors and growth factor RTKs have been implicated inthe abnormal proliferation and migration of pulmonary arterial smoothmuscle cells, including PDGF, EGF, FGF, and VEGF (Pullamsetti S. S.,PVRIReview (2009), 1(2): 124-128; Hassoun P. M., JACC Vol. 54, No. 1,Suppl S, (2009) S10-19.). Neointimal hyperplasia contributes toatherosclerosis, restenosis after percutaneous coronary intervention,and venous bypass graft disease. Vascular injury in each of theseconditions results in the release of mitogenic growth factors andhormones, which contribute to pathological vascular growth. Many ofthese molecules contribute to neointimal hyperplasia by activatingPI3-kinase in vascular smooth muscle cells (Sanada F., Circ. Res.(2009), 105; 667-675) and selective inhibition of PI3K-δ and -γ confersinteresting anti-inflammatory effects (Williams O., Chem. & Biol.(2010), 17:123-134). The protein serine/threonine kinases related toERK-1 and -2 transduce signals to the nucleus not in response to growthfactors and other mitogens but in response to cellular stresses such asinflammatory cytokines (IL-1β and TNFα). Ischemia kinases (JNKs) and p38likely play critical roles in the genetic response of many components ofthe cardiovascular system disease processes (Force T., CirculationResearch. (1996) 78:947-953) and have been suggested as targets forcardiovascular disease therapies (Force T., Circulation, (2004) 109(10):1196). The Rho-ROCK pathway also has an important role in mediatingvarious cardiac cell functions, including contraction, actincytoskeleton organization, cell adhesion and motility, proliferation,cytokinesis and gene expression, all of which are involved in thepathogenesis of cardiovascular disease. Abnormal activation of thispathway is associated with the pathogenesis of various cardiovasculardiseases such as hypertension, coronary and cerebral vasospasm,restenosis, atherosclerosis, stroke and heart failure, although theroles of the ROCK isoforms (ROCK1 and ROCK2) remain to be elucidated(Shimokawa H., Trends in Pharmacological Sciences, (2007),28(6):296-302).

Rheumatoid Arthritis (RA):

RA is characterized by leukocyte infiltration, synoviocyte hyperplasiaand osteoclastogenesis. Tyrosine kinases have key roles in the signalingpathways that regulate these processes (D'Aura Swanson C., Nat. Rev.Rheumatol. (2009) 5:317-324). Inhibition of receptor tyrosine kinases(RTK) such as platelet-derived growth factor receptors (PDGFR), vascularendothelial growth factor receptors (VEGFR) and Tie receptors have beenshown to reduce synovial hyperplasia and angiogenesis (Irvine K. M.,FASEB 20 (2006) E-1 to E-12). Non-RTKs are also important in RA. Forexample, signaling through Burton's tyrosine kinase results in B-celland T-cell activation while more-specific inhibitors of Janus kinasesand Syk, have already shown efficacy in the treatment of RA. Srcinhibition is expected to reduce monocyte maturation andosteoclastogenesis. In addition, blocking Kit activation may induce mastcell apoptosis, thereby reducing the production of inflammatorycytokines and degradative molecules in the synovium. The status ofcurrent approaches to kinase inhibitor based therapy for RA has beenreviewed recently (Muller S., Exprt. Opin. Drug Disco. (2010),5(9):867-881).

Discoidin Domain Receptors 1 and 2 (DDR1 and DDR2) are collagenreceptors with protein tyrosine kinase activity that control fundamentalcell processes including cell proliferation, adhesion, migration, andextracellular matrix remodeling (Vogel W, Cellular Signalling, 2006,18:1108-1116). These RTKs are important in embryonic development,skeletal growth, tissue repair and injury-induced remodeling of bloodvessels and the liver (Olaso E, J Clin Invest. 2001, 108:1369-1378;Zhang X H, Arch Med Res. 2010, 41(8):586-92; Ali B R, Hum Mol Gen, 2010,19(11):2239-2250; Hou G, Circ. Res 2002, 90:1147-1149; Ferri, N, Am JPathol, 2004, 164:1575-1585). Aberrant activity of both receptors hasbeen linked to human diseases such as lung, kidney and liver fibrosis,atherosclerosis, osteoarthritis, and rheumatoid arthritis. DDR1 and DDR2have also been implicated in primary and metastatic cancer progressionthrough regulation of metalloproteinase production, cell growth, andchemotactic invasion of normal tissue (Badiola I, Oncol Rep, 2011,26:971-978).

Hepatic fibrosis in response to chronic injury is similar in all formsof liver disease and involves type I collagen accumulation in thesubendothelial spaces between hepatocytes and endothelial cells. Thenewly generated fibrillar collagen replaces basement membrane likematrix containing type IV collagen. This conversion to fibrillarcollagen is pivotal in mediating the loss of differentiated functionthat characterizes progressive liver disease. Liver stellate cells arethe major source of fibrosis as they convert from quiescent cells toproliferative and fibrogenic myofibroblasts. In liver fibrosis mousemodels, DDR2 is upregulated in stellate cells following increasedcollagen synthesis and is an inducer of MMP-2 mediated growthstimulation suggesting this collagen receptor may help perpetuate thefibrosis (Olaso E, J Clin Invest. 2001, 108:1369-1378). DDR2 has alsobeen found at increased levels in the mesenchymal compartment as well asthe biliary epithelial cells in cirrhotic livers (Mao T K, Autoimmunity2002, 35(8):521.).

DDRs have also been shown to play a role in fibrosis of the kidney andlung. DDR1-null mice have also been found to have significantly reducedfibrotic and inflammatory responses in kidney hypertension models (VogelW, Cellular Signalling, 2006, 18:1108-1116.). The DDR1b isoform wasfound to be selectively induced in idiopathic pulmonary fibrosis (IPF)patients during disease progression and high levels of DDR1 can be foundin CD14 positive cells from bronchioalveolar lavage fluid from thesepatients compared to healthy volunteers or patients with other lungdiseases (Matsuyama W, FASEB J, 2003, 17(10):1286).

The discoidin domain receptors are also associated with inflammation andarthritis. LPS and IL-1β induces monocyte and neutrophil expression ofthe DDR1a and DDR1b isoforms. Transfection of DDR1a into leukemia celllines promotes adhesion while DDR1b enhances monocyte differentiation tomacrophages and upregulates their MIP-1α and MCP-1 production duringextravasation (Matsuyama W, J Immunol. 2005 174(10):6490). DDR1 isupregulated in activated T cells and can act as a co-stimulator undersuboptimal TCR/CD3 activating conditions (Dang N, J Immunother. 2009,32(8):773-784). The receptor kinase also enhances primary human T cellmigration through 3D collagen by a mechanism not dependent on adhesion(Hachehouche L N, Mol Immunol. 2010, 47(9):1866-1869; Chetoui N, J CellBiochem. 2011, 112(12)3666-3674).

DDR2 has been found to be integral in the maintenance and progression ofosteoarthritis and rheumatoid arthritis. DDR2 mediated MMP-13 inductionexacerbates the articular cartilage degeneration found in osteoarthritispatients. Reports both in mouse arthritis models and from human kneejoints found a correlation between increased DDR2 and MMP-13 expressionand the degree of type II collagen breakdown. These results suggest theperpetuation of DDR2 activation becomes a vicious circle where by DDR2promotes tissue catabolism which leads to cartilage damage and furtherDDR2 upregulation and activation (Sunk I G, Arthritis & Rheumatism,2007, 56(11):3685-3692.). Xu and his colleagues demonstrated thatreducing DDR2 expression by using DDR2^(−/+) heterozygous mutant miceled to decreased articular cartilage degeneration of the knee jointsinduced by injury or type XI collagen deficiency (Xu L, Arthritis &Rheumatism 2010, 62(9):2736-2744). These data suggest that regardless ofthe initiating event, osteoarthritis disease progression is perpetuatedby the continued activation of DDR2 and therefore therapeutic agentsthat specifically inhibit this kinase may be successful agents in theprevention and treatment of osteoarthritis.

The preponderance of evidence from research on discoidin domain receptorfunction demonstrates that DDRs are molecular sensors that monitorextracellular matrix integrity. However, aberrant or uncontrolled DDR1and DDR2 signaling has been associated with a variety of illnesses suchas arthritis, fibrotic disorders and cancer highlighting the potentialimportance of these collagen receptors in human health and disease.These data suggest DDR1 and DDR2 may be good targets for therapeuticintervention in multiple indications.

Pain:

Tropomyosin-related Kinases (Trk's) are the high affinity receptortyrosine kinases activated by a group of soluble growth factors calledneurotrophins (NT). There are 3 Trk receptor family members: TrkA, TrkBand TrkC. Trk's are widely expressed in neuronal tissue and areimportant in the maintenance, signaling and survival of neuronal cells(Patapoutian A., Current Opinion in Neurobiology, (2001), 11, 272-280).Inhibitors of the Trk/neurotrophin pathway have been shown to beeffective in many pre-clinical animal models of pain. For example,antagonistic NGF and TrkA antibodies have been shown to be efficaciousin inflammatory and neuropathic pain models (Woolf C. J., Neuroscience(1994), 62:327-331. Zahn P. K., J. Pain (2004), 5:157-163; Shelton D.L., Pain (2005), 116:8-16; Delafoy L., Pain (2003) 105:489-497;Theodosiou M, Pain (1999) 81:245-255; Li L., Mol. Cell. Neurosci.(2003), 23, 232-250; Gwak Y. S., Neurosci. Lett. (2003), 336: 117-120).Furthermore, several groups have demonstrated that BDNF levels and TrkBsignaling is increased in the dorsal root ganglion after inflammation(Cho H. J., Brain Res (1997) 764: 269-272.) and antibodies that decreasesignaling through the BDNF/TrkB pathway inhibit neuronalhypersensitization and the associated pain (Li C-Q., Molecular Pain,(2008), 4(28), 1-11).

It has also been reported that NGF secreted by tumor cells and tumorinvading macrophages directly stimulates TrkA located on peripheral painfibers. Using various tumor models in both mice and rats, it was shownthat neutralizing NGF antibodies inhibit cancer related pain to a degreeequal to or better than the highest tolerated dose of morphine. Inaddition, activation of the BDNF/TrkB pathway has been implicated as amodulator of neuropathic, inflammatory and surgical pain (Matayoshi, J.Physiol. (2005), 569:685-95; Thompson S. W. N., Proc. Natl. Acad. Sci.USA (1999), 96:7714-18; Li C-Q., Molecular Pain, (2008), 4(28), 1-11).These bodies of data suggest inhibitors of TrkA and/or other Trk kinasesmay provide an effective treatment for chronic pain states.

Kinases in Cancer:

Although kinase mediated pro-inflammatory or wound healing signalingpathways play important support roles in cancers (Karin M., Proc. Am.Thor. Soc. (2005) 2: 368-390; Rakoff-Nahoum S., Yale J. Biol. Med.(2006), 79:123-130; Gust T. C., Anti-Inflammatory & Anti-Allergy Agentsin Medicinal Chemistry, (2007), 6:19-27), these processes are usuallynot sufficient to initiate tumorigenesis. More often, the transformationprocess requires the aberrant activation of or activating mutation inkinases involved in other tumor specific key signaling pathways. Theterm ‘oncogenic addiction’ is often used when gene mutations provide asurvival advantage for tumor cells over non-transformed cells and theexpression of that gene product is required to avoid cell death. Theseoncogenes are commonly receptor tyrosine kinases (e.g. EGFR, PDGFRA,MET) or kinases in the PTEN/PI3K/AKT or Ras/Raf/MEK/ERK signalingpathways. For example, it has been estimated that 88% of allglioblastomas have altered signaling in one of these kinase pathways(Cancer Genome Atlas Research Network, Nature; (2008), 455:1061-1068).

The Ras/Raf/MEK/ERK and PI3K/Akt Pathways:

The RAS/RAF/MEK/ERK and the RAS/PI3K/PTEN/mTOR kinase cascades are twokey pathways that contribute to many cancers that are illustrative ofthe compensatory crosstalk and redundancies in signalling networks thatcan lead to development of innate or acquired resistance to theirindividually targeted therapies, FIG. 1 (Gibbony G. T. and Smalley K. S.M., Cancer Discovery (2013) 4(3): 260-263).

The Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) pathwaymediates cellular responses to different growth signals and isfrequently dysregulated in cancer. The RAF family proteins areserine/threonine specific kinases and are key players in the MAPKpathway. These proteins act immediately downstream of Ras to conductextracellular signals from the cell membrane to the nucleus via acascade of phosphorylation events. Thereby cell growth, proliferation,and differentiation can be regulated in response to growth factors,cytokines, and hormones (Christensen C., Oncogene (2005),24(41):6292-6302. Schnidar H., Cancer Res. (2009), 69(4):1284-1292.).The Ras/Raf/MEK/ERK pathway has been found to be upregulated inapproximately 30% of all cancers with higher percentages seen incutaneous melanomas as well as colon, lung, ovarian, and kidney tumors(Hoshino R. Oncogene, (1999) 18:813-822). Mutated RAS, especially KRAS,is seen in over 20% of all human cancers (Bamford S., Br. J. Cancer 91(2): 355-358; Bos J. L., A Review Cancer Res (1989) 49(17):4682-4689).RAS mutations have also been shown to lead to the promotion of PI3Ksignaling and dysregulation of the downstream RAF/MEK/ERK signalingpathways. Although early attempts to target Ras have not yielded anyviable drug candidates, many novel compounds inhibiting the activitiesof Raf and MEK have been developed and investigated in clinical trialsin recent years. Although the first MEK inhibitor (CI-1040) lackedefficacy in clinical trials, its low toxicity has encouraged the searchfor novel compounds with enhanced target potency (Wong K-K, RecentPatents on Anti-Cancer Drug Discovery, (2009), 4:28-35).

The three Raf kinases are designated as A-Raf, B-Raf, and C-Raf. At thistime only B-Raf (v-Raf murine sarcoma viral oncogene homologue B 1) isfrequently found mutated in various cancers (Palanisamy N., NatureMedicine (2010), 16(7):793-798.). The most common B-Raf mutationconstitutes 90% of all mutations to this kinase. The substitution of aglutamic acid residue for a valine moiety at codon 600 (V600E) resultsin a constitutively activated kinase that is ˜500-fold more active thanthe wild-type protein (Hoeflich K. P., Methods in Enzymology (2008),439: 25-38). This mutation, which occurs with a frequency of 50-70% incutaneous malignant melanoma, is also present in a wide range of otherhuman cancers, particularly thyroid (30%), colorectal (10%), and ovarian(35%) cancers (Flaherty K T, et al., NEJM (2010) 363:809-819; El-Osta H,et al., PLoS ONE (2011) 6(10):e258060). Advanced malignant melanoma hasa tendency to rapidly metastasize throughout the body and developresistance to treatment. In addition, melanoma rates continue to riseand the average patient age continues to decrease. Observation thatinhibition of B-Raf signaling blocks cancer cell proliferation andinduces apoptosis and its dysregulation in multiple tumor typesvalidates V600E B-Raf as an important therapeutic target with excellentopportunities for anticancer drug development. Increasedphosphatidylinositol 3-kinase (PI3K) signaling is also prevalent in manytypes of cancer (Vivanco I., Nat Rev Cancer (2002), 2(7):489-501; SerraV., Oncogene (2011) 2; 30(22):2547-57). Dysregulation of this pathwaymay be caused at a molecular level by activating mutations of PI3Kitself, by loss of PTEN, a negative regulator of PI3K activity,mutations in regulatory proteins, or by a variety of factors both up anddownstream of PI3K. PTEN is one of the most commonly mutated or deletedgenes in human cancer, second only to p53 (Cantley L. C., Proc Natl AcadSci USA. (1999) 96(8):4240-4245), and somatic mutations of the PI3Kp110α chain are found in 30% of all epithelial cancers (Engelman J. A.,Clin Cancer Res 2008; 14:2895-2899). In addition, P70S6K1, a kinasedownstream of PI3K/AKT pathway that is principle to the expression ofVEGF and survivin, has become a target of recent interest for cancertherapy (Skinner H. D., J. Boil. Chem. (2004) 279(44): 45643-45651; ZhaoP., Biochem. Biophys. Res. Commun. (2010), 395(2): 219-224). Finally,activation of the PI3K/AKT pathway has been strongly implicated inescape mechanism that compromise the effectiveness of specific kinasetargeted therapies (Wee S., Cancer Res. (2009) 69(10) 4286-4293; HynesN. E, Cancer Cell (2009)15: 353-355; Villanueva J., Cancer Cell (2010)15(6)):683-695; Paraiso K. H. T., Cancer Res. (2011) 71(7): 2750-60).

Stimulation of the Ras/PI3K/PTEN/AKT/mTOR pathway and hyper-activationof the Ras/Raf/Mek/Erk axis are dominant compensatory mechanism by whichinhibition of B-Raf(V600E) is ultimately circumvented (Davies M A etal., Cancer J. (2012) 18(2):142-7, Steelman L S, et al. J Cell Physiol.(2011) 226(11):2762-81; Yajima J, et al., Dermatology Research andPractice 2012; Article ID 354191), FIG. 1. Mutations that result in lossof PTEN function, activation of Ras, and/or loss of the RAS suppressingeffects of neurofibromin (via the NF1 gene) have been identified asmajor contributors to both the innate and acquired resistance to currentfront line B-Raf inhibitor therapies, FIG. 1 (Maertens O., Cancer.Discovery. (2013) 3(3); 338-49; Gibbony G T and Smalley K S M: 2013.

Other Important Receptor Kinases in Cancer:

Kinases upstream or outside of the Ras/Raf/Erk and PI3K/Akt pathway havealso been implicated in cancer cell differentiation and proliferationincluding the receptor tyrosine kinases in the Axl/Mer/TYRO3 and the Trkneurotrophin receptors (TrkA, TrkB TrkC) families.

The Axl/Mer/Tyro3 kinase family members have been implicated in tumorcell proliferation, cell-cell interactions, and cell migration andinvasion, suggesting multiple roles for this pathway in tumorigenesis.

Axl and Mer are expressed in various organs including the brain andtestes during development (Nagata K, J. Biol. Chem. (1996) 271 (47):30022-30027.). However in human adults their expression, which isnormally very low, returns to high levels in a variety of tumorsincluding glioblastoma, pancreatic, lung, thyroid, hepatocellular,colon, renal, gastric, and breast carcinomas (Funakoshi H., J. Neurosci.Res. (2002) 68:150-160; Li Y., Oncogene (2009), 28:3442-3455. ChallierC., Leukemia (1996) 10:781-787; Craven R. J., Int. J. Cancer (1995)60:791-797; Vajkoczy P., Proc. Nat. Acad. Sci. (2006):103(15):5799-5804; Sheih Y-S., Neoplasia. (2005) 7(12): 1058-1064; Xianzhou S.,Cancer (2011), 117(4):734-743). Tyro3 is also expressed in the brain andtestes and has also been linked to NK cell differentiation. RecentlyTyro3 has been identified as the upstream regulator ofmicrophthalmia-associated transcription factor (MITF), the ‘lineageaddiction’ oncogene in malignant melanoma. In animal models, blockingTyro3 repressed cellular proliferation and colony formation in melanomacells thereby inhibiting tumorigenesis in vivo (Zhu S., Proc. Nat. Acad.Sci. (2009) 106(4):17025-17030). Axl, Mer, and Tyro-3 mediate multipleoncogenic phenotypes and activation of these receptor tyrosine kinaseshas been shown to provide a mechanism of chemoresistance in a variety ofsolid tumors. The role of Axl and Gas6 in downstream signaling leadingto drug resistance involves a cancer cell's transition from anepithelial phenotype to one with mesenchymal properties(epithelial-to-mesenchymal transition or EMT). The EMT process allows acancer cell to acquire many of the hallmarks required for oncogenesisand drives the cell into a state that is more resistant to therapy. Theliterature suggests that selective inhibition of Axl signaling reversesEMT (Byer L A, et. al., Clin Cancer Res. (2013) 19(1):279-290) andshifts the cell back into a sensitive state which can then respond totargeted therapy. Targeted inhibition of these RTKs may be effective asanti-tumor and/or anti-metastatic therapy, particularly if combined withstandard cytotoxic therapies (Linger R. M. A., Targets (2010)14(10):1073-1090).

The Trk family of neurotrophin receptors are crucial for the normaldevelopment of the peripheral nervous system. These receptor tyrosinekinases signal through the PI3K, Ras/Raf/MEK and PLCγ1/PKC pathways andhave been found to play a critical role in neuroblastomas, the mostcommon and deadly solid tumor in children (Brodeur G. M., Clin CancerRes (2009) 15(10): 3244-3250). The Trk isoform expressed by theneruoblastoma can be prognostic as TrkA and TrkC expressing tumors aremore prone to spontaneous regression and a more favorable outcomewhereas TrkB are more often very aggressive and frequently haveconcomitant MYCN amplification. TrkB has been shown to suppress anoikis,or cell death induced by cell detachment, and thereby allowing themetastatic spread of tumor cells (Geiger T. R., Cancer Res (2007)67(13):6221-9). Trk family gene rearrangements or aberrant expressionhave also been identified in papillary thyroid carcinomas, breastcancers, non-small cell lung cancer, prostate cancer, pancreatic ductaladenocarcinoma, pediatric sarcomas, and leukemias (Tognon C., CancerCell (2002), 2:367-76; Liu Q., EMBO J (2000); 19: 1827-38; Eguchi M.,Blood (1999), 93:1355-63; Harada T., Clin Cancer Res. (2011),17(9):2638-45. Jones-Bolin S. E., Proc Amer Assoc Cancer Res (2005) 46:Abstract #3026).

Osteolytic metastases are common in many types of cancer and have beenfound in up to 70% of patients with advanced breast or prostate cancerand in approximately 15% to 30% of patients with lung, colon, stomach,bladder, uterus, rectum, thyroid, or kidney carcinomas. Bone metastasescan cause severe pain, hypercalcemia, pathologic fractures, spinal cordcompression, and other nerve-compression syndromes. Expression of TrkAand TrkC receptor kinases have been observed in the bone forming area inmouse fracture models and NGF expression was observed in almost all boneforming cells (Asaumi K., Bone (2000) 26(6): 625-633.). These datasupport exploring the use of pan Trk inhibitors for the treatment ofbone remodeling diseases such as bone metastases in cancer patients aswell as osteoporosis and rheumatoid arthritis.

Discoidin Domain Receptors 1 and 2 (DDR1 and DDR2) are collagenreceptors with protein tyrosine kinase activity that control fundamentalcell processes including cell proliferation, adhesion, migration, andextracellular matrix remodeling (Vogel W, Cellular Signalling, (2006)18:1108-1116). These RTKs are important in embryonic development,skeletal growth, tissue repair and injury-induced remodeling of bloodvessels and the liver (Olaso E, J Clin Invest. (2001) 108:1369-1378;Zhang X H, Arch Med Res. 2010, 41(8):586-92; Ali B R, Hum Mol Gen,(2010) 19(11):2239-2250; Hou G, Circ. Res (2002) 90:1147-1149; Ferri, N,Am J Pathol, (2004) 164:1575-1585). Aberrant activity of both receptorshas been linked to human diseases such as lung, kidney and liverfibrosis, atherosclerosis, osteoarthritis, and rheumatoid arthritis.DDR1 and DDR2 have also been implicated in primary and metastatic cancerprogression through regulation of metalloproteinase production, cellgrowth, and chemotactic invasion of normal tissue (Badiola I, Oncol Rep,(2011) 26:971-978).

DDR1 and DDR2 have been linked to several human cancers. DDR1 has beenfound in breast, ovarian, brain, esophageal, lung and immune systemcancers (Vogel W, Cellular Signalling, 2006, 18:1108-1116).). Barker etal demonstrated DDR1 was more highly expressed in cancerous breastepithelial cells than in adjacent normal breast tissue (Barker K T,gene, 1995 10:569). This collagen receptor has also been shown to be adirect transcriptional target for the p53 tumor suppressor gene. DDR1inhibition in tumor cells with wild type p53 activity results inincreased apoptosis (Ongusaha P P, EMBO J, (2003) 22(6): 1289). DDR2 hasbeen shown to play a role in breast, lung and immune system cancers aswell. Recently a group reported that approximately 3% of 277 lungsquamous cell carcinoma patients had DDR2 mutations (Kotz J, SciBX 2011,4(20):1-2). Though DDR1 and DDR2 have not been shown to be oncogenes incarcinogenesis, they likely act through regulating tumor cell growth,adhesion and metastasis by controlling collagenous extracellular matrixremodeling and metalloproteinase expression.

Other Non-Receptor Tyrosine Kinases in Cancer:

Several non-receptor tyrosine kinases such thirty-eight-negative kinase1 (Tnk1), JAK kinases, breast tumor kinase (Brk or PTK6), ROS, and ARGhave also been implicated in tumor progression, survival and metastasis.Though the tyrosine kinase Tnk1 has been identified as a tumorsuppressor gene in some cellular contexts, recently a novel genetranslocation has been identified that results in a fusion proteincombining part of C17ORF61 with Tnk1 kinase (Gu T-L., Leukemia (2010),24:861-865.). The TNK1-C17ORF61 fusion protein, which retainsconstitutive Tnk1 tyrosine kinase activity, was confirmed to drive theproliferation and survival of Hodgkin's lymphoma (HL) cell line, L-540.In addition, the application of functional genomics by using HT-RNAiscreens has allowed researchers to identify TNK1 as a growth-associatedkinase in pancreatic cancer cells (Henderson M. C., Mol Cancer Res.(2011) 9(6).).

Activating mutations in JAK family members are observed in leukemias andmyeoloproliferative neoplasms (Verstovsek S., Hematology (2009)636-642). Several lines of evidence support the conclusion that JAK/STATsignaling is exaggerated in hematological malignancies and likelycontributes to disease pathogenesis. Activating mutations in Jak1 havebeen described in acute lymphoblastic liekemia (ALL) and theJak2^(V617F) mutation is particularly important in myeleoproliferativeneoplasms (MPNs) and myelofibrosis.

Brk is a member of a novel family of soluble protein tyrosine kinases,considered to be distantly related to c-Src (Ostrander J. H., Cancer Res(2007); 67: 4199-4209). Brk has been shown to localize to the nucleus ofsome breast and prostate cancer cell lines and is coamplified andcoexpressed with ErbB2 in human breast cancers. Brk has been shown tointeract with EGFR and ErbB3 and the expression of Brk enhancesEGF-induced ErbB3 phosphorylation and the recruitment of p85phosphatidylinositol 3-kinase to ErbB3, which potentiates PI3K activity(Xiang B., Proc. Nat. Acad. Sci. (2008); 105(34): 12463-12468). Datafrom these recent studies place Brk in a novel signaling pathwaydownstream of ErbB receptors and upstream of Rac, p38 MAPK, and ERK5 andestablish the ErbB-Brk-Rac-p38 MAPK pathway as a critical mediator ofbreast cancer cell migration. Furthermore, overexpression of Brkconferred resistance to the ability of Lapatinib, an ErbB2 kinaseinhibitor, to inhibit ErbB2-induced proliferation.

ROS kinase is one of the last remaining orphan receptor tyrosine kinaseswith an as yet unidentified ligand and the normal function so thiskinase in different body tissues have not been fully identifies.However, ectopic expression, as well as the production of variablemutant forms has been reported in a number of cancers, such asglioblastoma mutifore and non-small cell lung cancer, suggesting a rolefor ROS kinase in deriving such tumors. The recent discovery of newselective inhibitors for ROS, along with the development of newdiagnostic tools for the detection of ROS fusion proteins, indicatesthat targeting of this kinase and its mutant forms may have clinicalapplications for the treatment of cancers (El-Deeb I. M. et al.,Medicinal Research Reviews, (2011) 31(5) 794-818).

ARG is an ABL-related kinase very similar to c-ABL at the SH3, SH2, andkinase domains and is expresses widely in normal cells (Krushe G. D.,Science (1986) 234:1545-1548; Perego R., Oncogene (1991) 6, 1899-1902).ARG is also implicated in leukemogenesis by the fusion between ARG andETV6 (ETS translocation variant 6), also known as TEL (translocation ETSleukemia). The ARG:ETV6 fusion was identified in two independent casesof human leukemias with t(1;12)(q25;p13) translocation (Cazzaniga G.,Blood, (1999). 94:4370-4373; Iijima Y. Blood, (2000) 95: 2126-2132) andin a T-lymphoblastic cell line derived from a patient with acutelymphoid leukemia carrying t(1;10;12)(q25;q23;p13) (Nishimura N.,Oncogene (2003) 22: 4074-4082). Constitutive activation of ARG and Ablkinases has also been implicated in the promotion of breast cancer cellinvasion (Srinivasan D., Cancer Res (2006) 66(11): 5648-55). It has beenreported that ARG is a target of the small molecule, tyrosine kinaseinhibitor STI571 (Okuda K., Blood (2001), 97:2440-2448) which maycontribute to the ability of STI571 (Imatinib/Gleevec) to inducehematologic remission in most patients with chronic myeloid leukemia.

Roles of Escape Mechanisms in the Treatment of Cancer:

Targeted inhibitors against specific tyrosine kinases known to becritical in tumor cell growth, differentiation, and survival havegenerated a lot of excitement over the last decade. Although there havebeen some dramatic examples of clinical responses in tumors known tohave genetic mutations in single genes, i.e. the BCR:Abl fusion proteinin CML and the B-Raf (V600E) mutation, highly specific kinase inhibitorscan be met ultimately with resistance and tumor escape due to pathwayenabling mutations in the target or suppressor proteins andup-regulation of compensatory proteins or pathways. In spite of recentadvances, improving the outcomes for patients afflicted with relapsedand refractory cancer still represents a significant challenge. Toooften, newly approved, targeted agents produce a significant upfrontresponse in cancer patients only to be followed by drug resistance andprogressive disease. Significant efforts have been made to understandthe mechanisms of drug resistance, particularly to targeted agents.

Tumor cells that harbor B-Raf(V600E) exhibit oncogenic addiction andtargeted inhibitors, such as the Type-I inhibitor Vemurafenib, havedemonstrated remarkable efficacy in advanced stage disease driven bythis mutation (Ribas A, et al., Clin Oncol (2011) 29:Suppl:8509; ChapmanP. B., et al. New Engl. J. Med. (2011) 364: 2507-2516]. However,resistance to Raf inhibitors, such as Vemurafenib, develops quickly(within 6-7 months) and recent studies have suggested that drugaddiction pays a role and that removal of drug may be required to haltthis life threatening resistance (Das M. et al. 2013 e-print,doi:10.1038/nature11814). In addition, cell population heterogeneity,compensatory pathway activation, inactivation of suppressor proteins,and external stimulation by the micro-environment can conspire topromote resistant disease, FIG. 1 (Gibbony G. T. and Smalley K. S. M.,Cancer Discovery (2013) 4(3): 260-263; Paraiso K H T, et al., ClinicalCancer Research, (2012) 18(9):2502-2514.).

Another limitation of Type-I, ATP competitive, B-Raf inhibitors have isdue to their ability to transactivate wild-type B-Raf and Raf-1 innormal cells (Hatzivassiliou G, et al Nature, (2010) 464:431-435;Heidorn S J, et al. Cell (2010) 140:209-221; Poulikakos P I, et al.Nature 2010 464:427-430) and intermittent treatment has again beenproposed as a means of improving patient outcomes (Thakur M D, et al.,Nature (2013) 494: 251-255). Therefore, efforts continue to understandthe limitations of current targeted therapies and escape mechanisms withimproved inhibitors and adjunct therapies.

Recently, new insights into the mechanisms of resistance have beenprovided (Maertens O. et al., Cancer Discovery (2012) 3(3): 338-349;Whittaker S. R. et al., Cancer Discovery (2012) 3(3): 350-362; Gibney G.T. and Smalley K. S. M., Cancer Discovery (2013) 4(3): 260-263).Although B-Raf mutations play a well established role in melanogenesis,without additional genetic alteration, tumor development is oftenrestricted to oncogene-induced senescence (OIS). Nf1 mutations suppressB-Raf induced senescence, promote melanocyte hyperproliferation, andenhance melanoma development. Nf1 mutations function by deregulatingboth PI3K and ERK pathways. As such, Nf1/B-Raf mutant tumors areresistant to B-Raf inhibitors but are sensitive to combined inhibitionof MAPK/ERK and mTOR. If Nf1 is mutated or suppressed in human melanomasthat harbor concurrent B-Raf mutations, the Nf1 ablation decreases thesensitivity of melanoma cell lines to B-Raf inhibitors. Importantly,loss of Nf1 activity is seen in patients following sustained treatmentwith B-Raf inhibitors and mechanisms of Nf1 inactivation have beenassociated with acquired or innate resistance to these targetedtherapies in melanoma.

Constitutive activation of signaling upstream or further downstream fromthe inhibited target protein is a common resistance mechanism. Forexample, blockade of mTOR with rapamycin analogs results in an increasein AKT signaling that reduces their overall therapeutic effect (ZitzmannK., Cancer Letters (2010), 295(1): 100-109; O'Reilly K. E., Cancer Res.(2006), 66: 1500-1508). In such cases, targeting multiple kinases in theaffected signaling pathway can maximize pathway inhibition.Consequently, mixed inhibitors of PI3K and mTOR have been developed(Brachmann S., Curr. Opin. Cell Biol. (2009) 21(2): 194-198; VenkatesanA. M., Bioorg. Med. Chem. Lett. (2010), 20(2): 653-656).

Another common cause of specific inhibitor resistance is through theactivation of a redundant receptor or parallel pathway that canfunctionally substitute for the inhibited one. This type of resistanceoccurs with receptor tyrosine kinases when related family members canperform overlapping functions and inhibiting one receptor cannotcompletely block downstream signaling. It has been found thatapproximately 20% of tumor samples from patients that became resistantto EGFR inhibitors had MET gene amplification (Engleman J. A., ClinCancer Res 2008; 14:2895-2899). The crosstalk between EGFR and MET,observed in breast cancer cells, explains the EGFR inhibitor resistanceseen in such tumors (Tao Y., Nat. Clin. Pract. Oncol, (2007) 4(10):591-602). Therefore an inhibitor that inhibits both EGFR and MET couldbe efficacious in treating such patients. Up-regulation of the PI3K/PTENsignaling through PIK3CA activating mutations or PTEN loss is anothermechanism found in EGFR inhibitor resistance (Janmaat J. L., Clin CancerRes, (2003), 9(6):2316-2326). Chemotherapy resistance has also been tiedto PI3K/Akt activation through EGFR (Winograd-Katz S., Oncogene (2006),25:7381-7390).

Of particular interest is the crosstalk between the PI3K/AKT/mTOR andRAS/Raf/MEK/ERK pathways often utilized by tumors as a compensatorymechanism when specific inhibitors of a single pathway are used (FaberA. C., Cell Cycle (2010) 9(5) 851-852). It has been shown that PI3Kinhibition in HER2-overexpressing breast cancers can lead to theup-regulation of the compensatory ERK signaling pathway. Inhibition ofboth PI3K and MEK simultaneously has been demonstrated to lead todecreased proliferation and superior anti-tumor activity in animalmodels and this combination therapy is currently being studied in theclinic (Worcester S., Elsevier Global Medical News. (2011) Apr. 11).

As described above, the issues surrounding the mono-specific TKIs hasled to the revitalization of interest in development of ‘dirtier’ kinaseinhibitors that hit multiple kinases at the same time. One earlysuccessful multikinase inhibitor is Sorafanib (Naxavar) which targetsRaf, VEGFR, PDGFR(3, FLT3, p38 and c-Kit all with IC₅₀s in the nanomolarrange. Other examples of approved multikinase inhibitors includeSunitinib (Sutent), Erlotinib (Tarceva) and Imatinib (Gleevac). However,these early versions of multikinases rarely hit both the Ras/Raf/ERK andPI3K/AKT/mTOR pathways at the same time.

It has been reported that tumors, such as AML acquire resistance tothese multikinase Raf/Flt3/c-Kit inhibitors, due to the activation ofcompensatory PI3K/AKT pathways after several months of treatment. It isbecoming increasingly apparent that inhibiting both the target oncogeneand kinases involved in the commonly used escape mechanisms will berequired to achieve durable responses with targeted cancer therapies.Single target TKIs currently in phase 1 or phase 2 clinical trials areproviding significant amounts of data on which pathways are commonlydysregulated in the most prevalent tumor types and more importantlywhich compensatory pathways lead to tumor escape. This information willhelp determine which specific inhibitors would be most effective givenin combination leading to more durable tumor growth inhibition in thepatients.

Another reoccurring mechanism that appears to be an underlying cause toboth upfront and acquired resistance to many receptor tyrosine kinase(RTK) inhibitors is the up-regulation of Axl. This has been demonstratedin multiple cancer types with numerous targeted agents, includingimatinib resistance in gastrointestinal stromal tumors (GIST), erlotinibresistance in non-small cell lung cancer (NSCLC) (Byers, et. al., ClinCancer Res. 2013; 19(1):279-290), PKC412 resistance in acute myeloidleukemia (AML), cetuximab resistance in squamous cell carcinoma of thehead and neck (SCCHN) (Giles, et al., Mol Cancer Res (2013)12(11):2541-2558), and lapatinib resistance in breast cancer (Liu L, etal., Cancer Res (2009) 69(17):6871-6878. Holland S, et. al. (Cancer Res.2010) 70:1544-1554). Axl is also one of the most common RTKs detected inbreast cancer (Meric F, et. al., Clin Cancer Res. (2002) 8:361-367)where expression promotes metastasis and is associated with a poorprognosis (Gjerdrum C, et. al., Proc Natl Acad Sci USA. (2010)107:1124-1129). Moreover, inhibition of Axl has been shown to restoresensitivity to targeted agents in a synergistic manner (Verma A, et.al., Mol Cancer Ther. (2011) 10(10); 1763-73). Axl is a member of theTAM receptor kinase family that includes Mer and Tyro3. The overexpression of any of the three family members has been associated withtumor cell survival and growth, increased migration, and angiogenesis(Linger R M, Adv Cancer Res. (2008) 100:35-83 and Linger R M., ExpertOpin Ther Targets. (2010) 14:1073-1090). AML cells are known to inducethe expression and secretion of the TAM receptor ligand Gas6 (growtharrest-specific gene 6) by bone marrow-derived stroma cells, which inturn mediates proliferation, survival and chemo-resistance in AML cells.Mer receptor tyrosine kinase over expression has been shown tocontribute to leukemogenesis (Lee-Sherick A B, Oncogene, (2013)32(46):5359-68) and its inhibition increases chemo-sensitivity anddecreases oncogenic potential in T-cell acute lymphoblastic leukemia(Brandao L N, Blood Cancer Journal, (2013) 3 (1): e101 DOI:10.1038/bcj.2012.46). A good case has also been made that Axl, Mer andTyro3 are potential targets in Melanoma and as an adjunct toimmunotherapies (Sensi M, et al, J. Invest. Derm. (2011) 131:2343-57;Schlegel J, et. al., J Clin Invest. (2013) 123(5):2257-2267; Demarest SJ, et. al., Biochemistry. (2013) 52(18):3102-18).

Deregulation of protein synthesis is also a common event in humancancers. A key regulator of translational control is elF4E and reportsindicate that eIF4E activity is a key determinant of bothRas/PI3K/Akt/mTOR and Ras/Raf/Mek/Erk mediated tumorigenic activity.Because activation of eIF4E involves phosphorylation of a key serine(Ser209) specifically by MAP kinase interacting ser/thr kinases (Mnk1and Mnk2) (Hou J., Oncotarget (2012) 3:118-131), efforts to discovereither selective or combined targeted inhibitors of these kinases areunderway by several research groups (Kassoum N, et. al.—2013). Indeed,Mnk inhibition by the antifungal agent Cercosporamide suppressesprimitive leukemic progenitors (CFU-L) from AML patients in adose-dependent manner (Altman J K, Blood (2013) 121(18) 3675-3681).Resistance in chronic myeloid leukemia (CML) is also facilitated throughelF4E over expression by blast-crisis granulocyte macrophage progenitors(GMPs) which then act as leukemia stem cells (LSCs) (Smith C C,Hematology Am Soc Hematol Educ Program. (2011) 2011:121-7). AlthoughelF4E activation is necessary for oncogenic transformation, it seemsdispensable for development of normal hematopoietic stem cells (HSCs).

Moreover, since Mnks act downstream of both MAPK and PI3K pathways,their inhibition may also have utility in Vemurafenib resistant cancerswhere Raf up-regulation and aberrant Ras/PI3K/Akt/mTOR axis activityconspire to promote resistant disease within 6-7 months (Davies M A,Cancer J. (2012)18(2):142-7). Simultaneous inhibition of both TAM andMnk family members should be more effective as an adjunct to targetedtherapies than either TAM or Mnk inhibition alone and data suggests thatsuch an agent may not significantly increase the side effect burden oftargeted therapies (Linger R M., Expert Opin Ther Targets. (2010)14:1073-1090).

Applications of Imaging Agents:

Use of imaging agents for monitoring disease progression is wellestablished (Smith-Jones K. M., J. Nuclear Medicine (1994), 35(2):219-325; Solit D. B, Cancer Res (2007), 67(23):11463-11469). Interesthas intensified regarding the application of such agents for thediagnosis, localization, and characterization of cancers (Hoffman J. M.,Radiology (2007) 244(1): 39-47; Stehouwer J. S., J. Med. Chem. (2010),53(15): 5549-5557) as well as both acute and chronic inflammatory anddegenerative diseases. More recently, applications directed specificallyat the monitoring of kinase activity have also been reported (Dumont R.A., Cancer Res. (2009), 69(7): 3173-3179; Samen E., Eur. J. Nucl. Med.Mol. Imaging (2009), 36:1283-1295; Pisaneschi F., Bioorg. Med. Chem.2010, 18: 6634-6645; Koehler L., European Journal of Medicinal Chemistry(2010) 45: 727-737.). The promise of imaging technologies for improvedbenefit, reduced cost, and personalization of medicine is significant.

General Construction of Kinase Inhibitors:

The general construction strategies and key structural elements forkinase inhibitors have been analyzed and reviewed extensively (Liu Y.,Nature Chemical Biology (2006) 2:358-364; Goshe A. K. J. Med. Chem.(2008), 51(17):5149-5171; Zhang J., Nat. Rev. Cancer (2009) 9(1): 28-39)and, based on their mechanism of inhibition, they can be classified aseither of five types, (Cozza G., Anti-Cancer Agents in MedicinalChemistry, 2009, 9:778-786).

Type-I inhibitors compete at the ATP binding site of a kinase andtypically bind to three subsites: 1) the purine binding site or “HingeRegion”, 2) the solvent exposed “Flap Region” at the entrance to the ATPsite, and 3) a lipophilic site adjacent to the purine site that is oftenreferred to as the “Gatekeeper Region”. Taken together, these bindingsites recognize “Hinge-Gatekeeper Motifs” (HGM) that can achieve usefulselectivity and profiles of inhibition, FIG. 2A. One major disadvantageof Type-I, ATP competitive, inhibitors is the kinetic challengeresulting from the millimolar physiological concentrations of ATP. Thesecond challenge is achieving selectivity for a particular kinase sincethe basic construction, functionality, and topography of ATP bindingsites are necessarily similar.

In contrast, Type-II inhibitors bind to an alternate inactiveconformation, exhibited by some kinases, in which a conservedAspartyl-Phenylalanyl-Glycine (DFG) containing loop is reoriented suchthat the Phenylalanine side chain is removed from its lipophilic bindingpocket, FIG. 2B. As a result of this conformational change, new bindingsites just adjacent to the “Gatekeeper Region” become accessible.Therefore, compounds that bridge from the HGM to these new “SelectivitySites” can build in new structural elements that take advantage ofdifferences between kinases in this region. Because the DFG-outconformation represents a minor population, Type-II inhibitors displaytime dependent kinetics resulting from slow on rates and the energeticsof conformational equilibration. Consequently, potent Type-II inhibitorsmust also exhibit slow rates of disassociation while further kineticadvantages of Type-II result from this conformations inability to bindATP and be recognized by upstream regulatory kinases. (Goshe A. K. J.Med. Chem. (2008), 51(17):5149-5171).

Typically, Type-II inhibitors incorporate a carboxamide, urea, orsimilar H-bond bridging bioisoster linkage to span from the SelectivitySites into the ATP binding region. Therefore, Type-II inhibitors can beviewed as a Hinge-Gatekeeper Motif (HGM) appropriately connected to alipophilic template that penetrates into and is complimentary to theadjacent “Selectivity Sites”. Using this construct, Type-I inhibitorshave been converted to Type-II inhibitors (Liu Y., Nature ChemicalBiology (2006) 2:358-364; Kufareva I., J, Med. Chem. (2008),51(24):7921-32).

Type-III inhibitors are relatively uncommon as they occupy a regionadjacent to but not overlapping with the ATP binding site that does notrequire significant conformational change of the DFG-loop. BecauseType-III inhibitors coexist with ATP binding, they are non-competitivewith ATP and unaffected by the high physiological ATP concentrations.Since Type-Ill inhibition is rarely observed, inhibitors of this typeoffer potential selectivity advantages over Type-I inhibitors. MEK1 andMEK2 are important kinases for which Type-III inhibitors have beenreported (Tecle H., Bioorg. Med. Chem. Lett. (2009), (19)1: 226-229).

Type IV inhibitors compete with a protein kinase substrates and targetregions outside the ATP binding site that may avoid some crucialproblems associated with the more conventional ATP competitive kinaseinhibitors, such as the development of drug resistance as a result ofaccumulating mutations in the ATP binding site of the kinase.

Type V inhibitors are defined as a family of allosteric inhibitors thatrecognizes a binding domain well outside the ATP-binding cleft and notnecessarily close to the substrate pocket. Therefore, this type ofinhibition can be very specific for a given kinase.

Use of 5-Membered Heterocyclic Scaffolds at the ATP Binding Site:

The tri-substituted imidazole template has been applied verysuccessfully to the Type-I inhibition of kinase. (Takle H., Bioorg. Med.Chem. Lett. (2009), (19)1: 226-229.) and knowledge gained from bindingat the ATP site (Bennett et. al., WO2007105058A2) has been extended byappending functionality to provide structurally related Type-IIinhibitors. (Tang J., Bioorg. Med. Chem. Letts., (2008), 18:4610-4614),FIGS. 3 A & B. In fact, structural information gained from Type-Iinhibitors has been translated to the design of a type-II inhibitorintentionally by the same group (Wolin R. L., Bioorg Med Chem Letts.(2008), 18(9):2825-2829), FIGS. 3 C & D. These examples illustrate howchemotypes that target the ATP binding site can be adapted to provideType-II inhibitors through an appropriate urea, amide, or ether, FIGS. 3E & F, linkage to an additional lipophilic aromatic ring that occupiesthe lower selectivity-site (Meyers M. J., Bioorg Med. Chem. Letts.(2010) 20:1543-1547). It is estimated that approximately 50 of the 518kinases adopt the DFG-out conformation (Fabian M. A., Nat Biotechnol(2005) 23:329-336), thus limiting the targets for Type-II inhibitors.However, recent studies suggest that the DFG-out conformation may bemore common than initially thought (Kufareva I, J, Med. Chem. (2008),51(24):7921-32).

Use of Urea-Linked 5-Membered Heterocyclic Scaffolds in Type-IIInhibitors:

Urea linked aryl-substituted 5-membered heteroaryl scaffolds have beenused previously to create favorable “Selectivity Site” interactions forthe Type-II inhibition of kinases, FIG. 4 A-D, (Smith R. A., Bioorg.Med. Chem. Letts. (2001), 11: 2775-2778; Regan J., J. Med. Chem. (2002),45: 2994-3008. Regan J., J. Med. Chem. (2003), 46: 4676-4686;Michellotti E. L., et. al., WO/2006/062982; Raeppel. S, Bioorg. Med.Chem. Letts. (2009), 19:1323-1328). The urea function acts as a criticalhydrogen bonding bridge between a conserved Glutamate side chain and theaspartyl NH from the DFG-loop. In addition to urea linages, carboxamidelinages have also been widely employed to profide Type-II inhibitors,(Zhang J, Nat. Rev. Cancer (2009) 9(1): 28-39). The closest prior art tothat described herein is the cyclic urea c-Met inhibitor illustrated inFIG. 4-D. To our knowledge, the only previous description of aType-IIkinase inhibitor containing an ortho-Aryl-substituted 5-memberedheteroaryl carboxamide scaffold is from our previous work, (Dietrich J.,Bioorg. Med. Chem. (2010), 18(1): 292-304), which is limited to theimidazole scaffold with a quinazolinone HGM, and is the only example ofa carboxamide linked aryl-substituted 5-membered heteroaryl Type-IIinhibitor. We have recently become aware of a recent patent (Son, J B,et. al., WO 2011093684) which claims the use of5-arylmethyl-2-methyl-pyrazole-4-carboxamide kinase inhibitors, seeTable 1, HGM#10. In this patent, the lack of direct aryl substitution onthe pyrazole ring significantly changes the scaffold geometry that isnecessary for the unique properties describe for the scaffolds herein.

Compared to the limited structural variations that have been utilized tointeract at the “Selectivity Site” of the DFG-out conformation ofkinases, the structural Hinge Gatekeeper-interacting Motif (HGM)variations that bind to the ATP binding site has been well studies. Asurvey of the kinase inhibitor are reveals this diversity, see Tables1-9. In these tables, the HGM amine is illustrated in the first columnwhile the amine capping group is illustrated, where applicable, in theadjacent column. Variations on the imatinib HGM are depicted in Table 1,while a series of difluoroanaline HGMs normally capped by a sulfonylgroup are depicted in Table 2. A series of biaryl HGM amines aresummarized in Table 3 and a series of HGMs with Bicyclic GatekeeperInteracting Ring Systems are illustrated in Table 4. Following Tables 5and 6, which illustrate the wide variety of diaryl ether HGMs that havebeen explored, Table 7 displays related but atypical linker strategiesbetween the Hinge and Gatekeeper-interacting groups. Table 8 illustrateshow derivatives of heteroaryl linked ATP site inhibitors can be adaptedto the design of Type-II inhibitors. Lastly, Table 9 illustratednon-amine HGMs that, although not directly applicable to theconstruction of Type-II inhibitors, could be adapted, as indicated, forconstruction of Type-II inhibitors.

TABLE 1 Hinge Gatekeeper-interacting Motifs (HGMs) with 4-methyl-1-3Disubstituted-Phenyl Amine Gatekeeper-interacting Groups. ATP-SiteInterating Group-Hinge- Type-II-Selectivity Site HGM # GatekeeperMotif-HGM Interacting Group 1

2

3

4

5

6

7

8

9

10

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 1 Imatinib ABL Zimmerman 19961IEP.pdb KDR, Kit, Zimmermann, J. et al. Bioorg. Imatinib in ABL PDGFR,Med. Chem. Lett. 1996, 6, CSF-1R, 1221-1226. Order# A2308 FLT-3, DDRsales@tciamerica.com 2 Zhang 2010 WO2010-US41872 3 Masitinib KitA01.743.786 (AB1010) PDGFR www.aurorafinechemicals.com FGFR3 4 PonatinibBCR-ABL, Huang 2010 3CS9.pdb mutants, Huang, WS et al. Journal ofPoltinib in ABL KDR, FGFR1, Medical Chemistry 2010, 53 PDGFRα, (12):4701-19 FLT3, LYN 5 GSK Cpd- CSF-1R Baldwin 2008 Best CSF-1R 14d LCK,EGFR, Baldwin I., etal. Bioorg. Med. from libratry ErbB4, KDR Chem.Lett. 18 (2008) 5285- approach 5289 6 Bafetinib ABL LYN Kamitsuji 2007(INNO-406) Kamitsuji Y, et al. Bioorg Med Chem Lett. (2007) 17: 12-17 7AZ628 B-RAF Aquila 2007 PDGFR-a/b Aquila, B., Lyne, P., Pontz, T.: KIT,CSF1R WO2007113558 (2007) 8 TIE-2 Hodous 2007 Hodous B. L., et.al.Bioorg. Med. Chem. Let. (2007) 17: 2886-1889 9 ZM 336372 CRAFHall-Jackson 1999 Hall-Jackson C. A., et.al. Chem Biol (1999) 6(8): 559-68 10 1318242-17- Son 2011 9P Son, J B, et.al., WO 2011093684

TABLE 2 Representative 4-6-Difluoro-1-3-disubstituted GatekeeperContaining Inhibitors. ATP-Site Interating Reference or Comment or HGMGroup-Hinge- Inhibitor Kinases Vendor Crystal # Gatekeeper Motif-HGMSulfonamide Cap Name or ID# Inhibited Source Structure 11

B-RAF Ignacuo 2011 Ignacio A., et.al. WO 2011025940 Novel Type-IInhibitor 12

vemurafenib (Zelboraf) PLX-4032 B-RAF (V600E) 31 nM C-RAF (48 nM) SRMS(18 nM) MAP4K5 51 nM FGR (63 nM) B-RAF 100 nM Bollag 2010 Bollag, G.,et.al., Nature (2010) 467: 596-599 3OG7_B.pdb Raf (V600E) 13

PLX-4720 B-RAF Tsai 2008 Tsai, J., et.al., Proc. Natl. Acad. Sci. Usa(2008) 105: 3041-3046 3C4C.pdb B-RAF 14

PLX-3203 B-RAF Tsai 2008 Tsai, J., et.al., Proc. Natl. Acad. Sci. Usa(2008) 105: 3041-3046 3C4D.pdb BRAF 15

B-RAF Wenglowsky 2011 Wenglowsky, S., et.al. ACS Medicinal ChemistryLetters (2011) 2: 342-347 3TV4.pdb BRAF 16

B-RAF Wenglowsky 2011 Wenglowsky S., et.al., Bioorg. Med. Chem. Lett.(2011) 21: 5533-5537 3TV6.pdb BRAF 17

B-RAF Joachim 2011 Joachim R., et.al., WO(2011) 025951 A1 Novel Type-IInhibitor 18

Wenglowsky 2011 Wenglowsky S., et.al., Bioorg. Med. Chem. Lett. (2012)22: 912-915 Novel Type-I Inhibitor 19

B-RAF VEGFR-2 Ren 2012 Ren, L., et.al. Bioorg. Med. Chem. Lett (2012).22: 3387-3391 4E4X.pdb B-Raf

TABLE 3 Representative Biaryl-amine Hinge Gatekeeper-Interacting Motifs(HGMs). ATP-Site Interating Group-Hinge- Type-II-Selectivity Site HGM #Gatekeeper Motif-HGM Interacting Group 20

21

22

23

24

25

26

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 20 AC-220 FLT-3 WO/2005/048953;WO/2009/038757 21 BRAF Berger 2009 3II5.pdb Berger, D. M., et.al. BRAFBioorg. Med. Chem. Lett. (2009) 19: 6519-6523 US20070219186 22PF-4594755 PYK2 Shena 2011 Shena C. J., et.al., Experimental CellResearch (2011) 317: 1860-1871 23 Linifanib PDGFR-b Shankar 2007(ABT-869) CSF-1R D. B. Shankar, etal. BLOOD 375.4 KDR 109(8), 2007,3400-3408 WO/2004/113304 24 KDR, TIE-2 Dai 2008 Dai Y., et.al., Bioorg.Med. Chem. Lett. (2008) 18: 386- 390 25 KDR Dai 20085 Dai Y., et.al., J.Med. Chem. (2005) 48: 6066-6083 26 KDR, TIE-2 Miyazaki 2005 Miyazaki Y.,et.al., Bioorg. Med. Chem. Lett. (2005) 15: 2203-2207.

TABLE 4 Representative HGMs with Bicyclic Gatekeeper Interacting RingSystems. ATP-Site Interating Group-Hinge- Type-II-Selectivity Site HGM #Gatekeeper Motif-HGM Interacting Group 27

28

29

30

31

32

33

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 27 Chen 2007 Chen, Ning; Hu, EssaWO 2007-US16383 28 BRAF Smith 2009 3IDP.pdb Smith, A. L., et.al., BRAFJ. Med. Chem. (2009) 52: 6189-6192 29 Raf-265 RAF Amiri P, et al.(CHIR-265) VEGFR US20070299039 Mol Cancer Ther 2010; 9: 358- 368 30Raf265 RAF Amiri P, et al. des-methyl VEGFR US20070299039 derivative 31RAF US 2003-675927 VEGFR WO 2003-US10117 32 KDR Hasegawa 2007 Hasegawa,M., et.al., J. Med. Chem (2007) 50: 4453- 4470 33 Bauer, D., et.al.,Bioorg. Med. Chem. Lett. (2008) 18: 4844-4848

TABLE 5 Representative Diaryl Ether Hinge Gatekeeper-Interacting MotifsHGMs. ATP-Site Interating Group-Hinge- Type-II-Selectivity SiteInhibitor HGM # Gatekeeper Motif-HGM Interacting Group Name or ID# 34

BIRB 796 Dorama- pimod 35

Sorafenib Nexavar 36

Regorafenib BAY 73-4506 37

38

39

40

41

Comment or Kinases Crystal HGM # Inhibited Reference or Vendor SourceStructure 34 p-38 Regan 2003 IKV2.pdb Regan, J., et.al. J. Med. Humanp38 Chem. (2003) 23; 46(22): 4676- MAP Kinase in 86. Complex with BIRB796 35 KDR, Kit, Wood 1998 PDGFRs, Reidl B., et.al., CSF-1R,WO-1998-53559 FLT-3, DDR, Raf, Tie2 36 VEGFR-2/3, Onyx-Sorafenib RET,KIT, U.S. Pat. No. 7,351,834 (filed on 1999) PDGFR, and Rafs 37 B-RafNiculescu-Duvaz 2009 Niculescu-Duvaz, D., et.al., J. Med. Chem. (2009)52: 2255-2264 38 B-Raf Menard 2009 Menard, D., et.al., J. Med. Chem.(2009) 52: 3881-3891 39 B-RAF Okaniwa 2012 4DBN.pdb VEGFR-2 Okaniwa, M.,et.al., BRAF J. Med. Chem (2012) 55: 3452- 3478 40 KDR TIE-2 Hasegawa2007 Hasegawa, M., et.al., J. Med. Chem (2007) 50: 4453- 4470 41 B-RAF_((V600E)) Whittaker 2010 BRAF, CRAF, Whittaker S., et.al. Cancer SRC,LCK, Res; (2010) 70(20) 8036- PDGFR-a, p38- 8044. a, p38-g,

TABLE 6 Representative Diaryl Ether Hinge Gatekeeper-Interacting MotifsHGMs continued. ATP-Site Interating Group-Hinge- Type-II-SelectivitySite HGM # Gatekeeper Motif-HGM Interacting Group 42

43

44

45

46

47

48

49

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 42 KI-20227 CSF-1R > Ohno 2008 Rand S isomers 480.54 KDR > Kit, Ohno, H. et al. Eur. J. are slightlyPDGFR-b Immunol. (2008) 38: 283-291. different 43 BMS 777607 c-METSchroeder_2009 Schroeder 3F82.pdb GM, et.al. J. Med. Chem. (2009) 52(5):1251-1254. 44 c-MET Schroeder 2009 3CE3.pdb Schroeder G. M., et.al. J.Med. Chem. 52(5), 2009, 1251-4 45 c-MET Schroeder_2009 Schroeder3CTH.pdb GM, et.al. J. Med. Chem. (2009) 52(5): 1251-1254. 46 Foretinibc-MET Qian 2009 GSK1363089 VEGFR-1, Qian F., et.al., Cancer Res XL880VEGFR-2 (2009) 69: 8009-8016 EXEL-2880 47 MGCD265 c-MET U.S. Pat. No.VEGFR-1, 7,772,247 VEGFR-2, VEGFR-3, RON and TIE2 48 Lenvatinib VEGFR2/3 Matsui 2008 (E7080) Matsui, J.; et.al., Clinical Cancer Research(2008) 14 (17): 5459-65 49 PYP-4-0001 Scientific Laboratory Inc.sales@sphinxscientificlab.com

TABLE 7 Hinge Gatekeeper-Interacting Motifs (HGMs) with Atypical LinkerGroups. ATP-Site Interating Group-Hinge- Type-II-Selectivity Site HGM #Gatekeeper Motif-HGM Interacting Group 50

51

52

53

54

55

56

57

58

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 50 Astra CSF-1R> D. A. Scott etal.Bioorg Med. Zeneca EphA2, Hck, Chem. 198(2008) 4794-4797 Cpd-27 Fyn,cRaf, WO/2007/071955 KDR, Src PDGFRs 51 KIT CSF1R US20070032519WO/2007/013896 52 p38 Milian 2011 2YIX.pdb Millan, D. S., et.al., p38and J. Med. Chem. (2011) 54: 7797 trialopyridine inhibitor 53 PF-4618433PYK2 Han 2009 3FZT[.pdb PYK2 Han, S., et.al., J. Biol. Chem. andPF-4618433 (2009) 284: 13193-13201 54 Cytopia- CSF-1R C. J. Burns etal.Bioorg. Med. Stereo Australia Kit, DDR1 Chem. Lett. 19 (2009) 1206-Chemistry 441.48 PDGFRs, 1209 Important VEGFR-1,2,3 FRK, RET 55Telatinib PDGFR-b Neeltje 2009 (BAY 57- VEGFR-2/3 Neeltje Steeghs N., JClin 9352) KIT Oncol (2009) 27: 4169-4176 56 RAF Bartkovitz D. J., etal., US20070060607 57 MK-2461 c-Met Pan B-S., et.al., Cancer Res Ron,Flt1-4, 2010; 70(4), 1524-1533 PDGFRβ, FGFR1-3 KDR, TrkA/B and Mer 58PYP-4-0025 Scientific Laboratory Inc. sales@sphinxscientificlab.com

TABLE 8 Hinge Gatekeeper-Interacting Motifs (HGMs) with Azole LinkingRing Systems. ATP-Site Interating Group-Hinge- Type-II-Selectivity SiteHGM # Gatekeeper Motif-HGM Interacting Group 59

60

61

62

63

64

Comment or Inhibitor Kinases Crystal HGM # Name or ID# InhibitedReference or Vendor Source Structure 59 Dabrafenib BRAF Stellwagen J.C., et.al. Bioorg. GSK2118436A Med. Chem. Lett. (2011) 21: 4436-4440WO201104441 WO2011044414 WO2011047238 60 BRAF Tang 2008 Tang J., et.al.,Bioorg. Med. Chem. Lett. (2008) 18: 4610- 4614 61 TIE-2 Lee 2010 Lee J.,et.al., Bioorg. Med. Chem. Lett. (2010) 20: 1573-1577 62 TIE-2 AdjabengG., etal. WO 2009076140 A1 63 B-RAF Stellwagen J. C. etal., Bioorg. Med.Chem. Lett. (2011) 21: 4436-4440 WO2009032667 A1 64 BKM120 Pan-PI3KBurger 2011 NVP- Burger T. M., et.al., ACS BKM120 Med. Chem. Lett.(2011) 2: 774-779

The structures in Table 9 represent kinase inhibitors that containinteresting HGMs that, because they lack an appropriately positionedamine group, can not be directly used to construct Type-II inhibitors.As illustrated in FIG. 3, ATP binding site inhibitor motifs from Type-Iinhibitors have been adapted to the construction of Type-II inhibitors(Wolin R. L., Bioorg. Med. Chem. (2010), 18(1): 292-304; Meyers M. J.,Bioorg Med. Chem. Letts. (2010) 20:1543-1547).

TABLE 9 Type-I Inhibitors that could be adapted to Library HGM Amines asIndicated. Type-II- Select- ivity Site Reference Comment ATP-SiteInterating Inter- Inhibitor or or HGM Group-Hinge- acting Name orKinases Vendor Crystal # Gatekeeper Motif-HGM Group ID # InhibitedSource Structure 65

Crizotinib PF02341066 ALK 66

None- Type-I Dasatinib ABL KDR, Kit, PDGFR, CSF-1R, FLT-3, DDR Lombardo,L. J. et al. J. Med. Chem. 2004, 47, 6658- 6661. 2GQG.pdb Dasatinib inABL 67

Want Reverse Amide Geuns-Meyer, S. D.; et. al., WO 2005113494 A2 68

Want Reverse Amide PCT Int. Appl., 2005113494, 01 Dec. 2005 69

Replace Benzyl- oxy Group with Amine Function Pfizer-12b 448.47 CSF-1RMeyers 2010 Meyers M. J., et al. Bioorg. Med. Chem. Lett. (2010)20:1543-1547 3LCO.pdb CSF1R 70

Replace Meth- oxy Group with Amine Function Pl3K/ mTOR Liu 2010 Liu, K.K.-C., et. al., Bioorg. Med. Chem. Lett. 2010, 20 (20), 6096- 6099.3ML9.pdb Pl3K- gamma 71

Replace Hydroxy Group with Amine Function WO 2007- 123892 72

None- Type-I Add Amine Functions as Indiacated PD173955 ABL KIT, SRC,HCK, LCK, Klutchko 1998 Klutchko, S. R. et al. J. Med. Chem. 1998, 41,3276- 3292. 1M52.pdb Abl 73

Pelitinib (EKB- 569) (EGFR) ErbB-1, −2 and −4 J. Med. Chem. 2003;46,49-63 Covalent Inhibitor

An efficient synthesis of2-trifluoromethyl-4-aryl-imidazole-5-carboxylic acids FIG. 5A, has beenreported (Hagiwara K., et. al, WO/1995/004724). Quinazolinone containingamides, FIG. 5B, have been claimed to potently inhibit the oncongenicB-Raf (V600E) mutant kinase (Aquila B., et. al, WO2006/024834).Attachment of a 2-trifluoromethyl-4-aryl-imidazole-5-carboxylic acid, A,to this quinazolinone containing (HGM) resulted in a homologous seriesof hybrid compounds, FIG. 5C, some of which were reported to be highlypotent B-Raf(V600E) inhibitors (Dietrich J., et al. Bioorg. Med. Chem.(2010), 18(1): 292-304).

This previous disclosure described computational experiments using1UWJ.pdb, the co-crystal structure of B-Raf(V600E) with Sorafenib, inwhich no attractive low-energy pose were produced during dockingstudies. This poor fit, thought to result from a steric clash betweenthe inhibitor and Glutamate 500 in 1UWJ.pdb, FIG. 6A, promptedexploration of alternate binding modes. Thus, this report proposed analternative mode of binding based on docking experiments using theDFG-out monomer-B co-crystal structure of B-Raf(V600E) with PLX-4720(Tsai J., PNAS USA. (2008) 105(8):3041-6). By utilizing this co-crystalstructure, 3C4C.pdb, the proposed negative steric interaction withglutamate 500 appeared to be removed, FIG. 6B.

FIG. 6 illustrates the binding mode of quinazolinone containinginhibitors with B-Raf.

This publication also described the structure activity relationshipwithin a small series of structurally related compound which all sharedthe quinazolinone HGM and imidazole scaffold. The only structuralvariations reported were limited to the 2- and 5-positions of animidazole scaffold with the three compounds depicted in FIG. 7displaying the most potent activity against B-Raf(V600E). The apparenttime dependence reported was consistent with Type-II inhibition. Thephosphor-protein gel assay used in this report revealed sub-nanomolarIC50 values for these three inhibitors when the inhibitor and enzymewere preincubated for 1 hour prior to addition of the MEK1 substrate.When evaluated against a panel of 96 kinases, Table-10, all threecompounds displayed potent and selective inhibition of B-Raf(V600E),B-Raf(Wt), C-RAF(Raf-1), PDGFR-α, PDGFR-β, c-Kit, and p38-α.

TABLE 10 Summary of inhibition profiles for the imidazole quinazolinoneinhibitors KIN-035, KIN-038, and KIN-057, (Dietrich, 2010). InhibitorCode KIN-035 KIN-038 KIN-057 Ambit Biosciences Gene Symbol

ABL1 ABL1(E255K) Inhibition Scale ABL1(T315I)    X > 25% @ 1 μM ACVR1B   XX > 50% @ 1 μM ADCK3    XXX > 75% @ 1 μM AKT1 XXXX > 90% @ 1 μM AKT2ALK AURKA AURKB AXL BMPR2 X BRAF XXXX XXX XXX BRAF(V600E) XXXX XXXX XXXBTK X CDK11 XX X XX CDK2 CDK3 X CDK7 X CDK9 CHEK1 CSF1R XXX X CSNK1DCSNK1G2 DCAMKL1 X X X DYRK1B EGFR X EGFR(L858R) X EPHA2 XX ERBB2 X XERBB4 X ERK1 FAK FGFR2 FGFR3 X FLT3 GSK3B IGF1R IKK-alpha X IKK-beta XINSR JAK2 X (catalytic) JAK3 (catalytic) JNK1 JNK2 JNK3 KIT XXXX XXXXXXX KIT(D816V) X LKB1 MAP3K4 Inhibition Scale MAPKAPK2    X > 25% @ 1 μMMARK3    XX > 50% @ 1 μM MEK1    XXX > 75% @ 1 μM MEK2 XXXX > 90% @ 1 μMMET MKNK1 XX X MKNK2 X MLK1 p38-alpha XXXX XX XXX p38-beta X X X PAK1PAK2 PAK4 PCTK1 X PDGFRA XX XX X PDGFRB XXX XX XXX PDPK1 PIK3C2B PIK3CAX X PIK3CG PIM1 PIM2 PIM3 PKAC-alpha PLK1 PLK3 PLK4 X X X PRKCE X X XRAF1 XXXX XXXX XXXX RET RIOK2 X X ROCK2 RPS6KA3 (Kin.) SNARK SRC SRPK3TGFBR1 TIE2 TRKA TSSK1B TYK2 X X (catalytic) ULK2 X X VEGFR2 X YANK3ZAP70 Compounds were assayed at 1 μM concentrations in duplicate withoutpreincubation. An X-indicates better than 25% inhibition, XX-indicatesbetter than 50% inhibition, XXX-represents better than 75% and XXXXrepresents better than 90% inhibition under the conditions of theseassays. Note atypical Structure-Activity Relationship of KIN-035 forCSF1R compared to KIN-38 and KIN-57.

The present disclosure addresses and interconnects two importantapplications relating to the treatment of diseases. Becauseunderstanding of the disease specific roles, complex interactions,mechanisms of dysregulation, activating mutations, and compensatoryback-up systems of kinase pathways, i.e. “Systems Biology”, continues toevolve, multi-targeted kinase (MTK) inhibitors with unique propertiesand selectivity profiles will continue to be needed. The development ofadvanced tools for the non-invasive mechanism-based characterization andmonitoring of disease in preclinical, clinical, and therapeutic settingsis also perceived as a critical unmet need.

The present disclosure describes a novel scaffold geometry and itsapplication to the design and preparation of selective or multi-targetedkinase (MTK) inhibitors as therapeutic agents and/or disease specificPET imaging agents. Enabling technologies for the early diagnosis,accurate characterization, patient specific treatment, and real timemonitoring of therapies will be essential for the realization ofpersonalized medicine. By combining therapeutic and imaging agents thatshare similar structural elements and/or activity profiles, significantsynergies can be realized in clinical development and personalizedmedicine. Taken together, these concepts constitute a platformtechnology with unique applications and utilities. Although thisdisclosure focuses on representative examples for the purposes ofillustration, the implications of and applications for this platformtechnology are quite broad and, using the information disclosed herein,one skilled in the construction of Type-I and/or Type-II kinaseinhibitors and the mode of binding of Hinge-Gatekeeper interactingMotifs (HGM) should be able to easily employ this platform technology.

SUMMARY

The present disclosure describes compounds of Formula (I):

or a pharmaceutically acceptable salt thereof, wherein the values forAr, A, B, D, E, R¹, G, L, and (H) are as defined herein.

Provided compounds and pharmaceutical compositions thereof are novelkinase inhibitors and are useful in treating a variety of diseases andconditions.

The present disclosure also provides the application of the combinationof functionality and scaffold geometry defined by Formula (I), or apharmaceutically acceptable salt thereof, to the design of novel kinaseinhibitors as imaging agents. Appropriate selection of the substitutedheterocyclic ring, R¹, Ar, (H)-L-G- and A/B/D/E can provide a variety ofkinase inhibitors with diverse and often unique selectivity profiles.This specific scaffold geometry is uniquely complementary to the DFG-outconformation of many kinases yet tightly anchors the inhibitor so thatinteractions with a particular kinase, or profile of kinases, can beselectively enhanced.

The disclosure also provides a pharmaceutical composition comprising adisclosed kinase inhibitor, including a compound of Formula (I), and apharmaceutically acceptable carrier or diluent, wherein the values forthe variables are as described below for the compounds of Formula (I).

The disclosure further provides a method of inhibiting kinases,comprising administering to a mammal in need thereof an effective amountof a disclosed kinase inhibitor, including a compound of Formula (I),wherein the values for the variables are as described below.

The disclosure further provides a method of imaging tumors with theB-Raf(V600E) mutation, comprising administering to a mammal in needthereof an effective amount of a disclosed kinase inhibitor, including acompound of Formula (I), wherein the values for the variables are asdescribed herein.

This disclosure further provides therapeutic agents and imaging agentswith unique selectivity profiles useful for the treatment of variouscancers resulting from the appropriate selection of the substituted5-membered heterocyclic scaffold, (H)-L-G-, Ar—, and A/B/D/E.

Also included in the present disclosure is the use of a disclosed kinaseinhibitor, including a compound of Formula (I), or a pharmaceuticallyacceptable salt thereof, for the manufacture of a medicament forinhibiting kinase activity in a mammal in need of such treatment.

Also included in the present disclosure is the use of a disclosed kinaseinhibitor, including a compound of Formula (I), or a pharmaceuticallyacceptable salt thereof, for the manufacture of a medicament fortreating a disease or disorder related to the activity or expression ofkinases, or for diseases or disorders in which activating mutations in,over expression of, or aberrant activity of kinase targets contributesto progression of a disease or disorder.

Also included in the present invention is a pharmaceutical compositioncomprising a disclosed kinase inhibitor, including a compound of Formula(I), or a pharmaceutically acceptable salt thereof, and apharmaceutically acceptable carrier or diluent.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Illustrates how proliferation can be stimulated through eitherthe Ras/Raf/Mek/Erk or Ras/PI3K/Akt/mTOR pathways and how loss of theRas negative regulatory protein Nf1 can enhance proliferation andcontribute to resistance to targeted PI3K or MAPK inhibitor therapies.

FIG. 2: Illustrates the conformational change for B-Raf (V600E) betweenA) DFG-In and B) DFG-Out populations and where Type-I, Type-II, andType-III inhibitors interact.

FIG. 3: Illustrates representative trisubstituted imidazoles A) and C)known to interact at the Hinge and Gatekeeper Sites that have beenappended to provide type-II inhibitors B) and D) as well as the strategyuse to convert Type-I inhibitor C) to Type-II inhibitor.

FIG. 4: Illustrates urea linked aryl-substituted 5-membered heteroarylkinase inhibitors.

FIG. 5: Proposed retrosynthesis of disclosed imidazole containing B-Rafinhibitors.

FIG. 6: Illustrates the published binding mode of quinazolinonecontaining inhibitors with B-Raf.

FIG. 7: Shows 2-CF3-imidazole quinazolinone inhibitors previouslyprofiled (Deitrich, et al. 2010).

FIG. 8: Illustrates the differences between the previously-describedB-Raf binding mode hypothesis for KIN-035. (Panel-A) and the new mode ofbinding (Panel-B) described herein.

FIG. 9: Graphic of 4JVG.pdb (wild type B-Raf) and 1UWJ.pdb (B-Raf(V600E)mutant and the corresponding cartoon drawings that illustrate thedifferent positioning of Lys⁶⁰¹ in these two structures.

FIG. 10: A graph of real-time kinetic data for SFE-0013 withB-Raf(V600E) that illustrate kinetics consistent with extremely tight orirreversible inhibition.

FIG. 11: Panel A) SFE-in B-Raf(V600E) with Lys601 moved slightly tobetter interact with or displace the 2-MeSO₂-imidazole substituent.Panel B) Putative covalent adduct from reaction with Lys601 (yellow) vsoriginal Lys601 position

FIG. 12: Summary of SAR studies for imidazole quinazolinone inhibitors.

FIG. 13: Represents inhibitors modeled into the crystal structures ofB-RAf, p38-α, CSF1R, KDR, FLT-3, and c-Met.

FIG. 14: Illustrates Hinge-Gatekeeper-Selectivity Site interactions forc-Met inhibitors.

FIG. 15: Illustrates certain inhibitors and affinity probes for theidentification of new atypical kinase conformations.

FIG. 16: Represents an exemplary method for the incorporation of ¹⁸F or¹¹C labels for PET probe preparation.

FIG. 17: Illustrates a strategic route for [18]F incorporation with(1,3-Bis(2,6-diisopropylphenyl)-2,2-difluoro-2,3-dihydro-1H-imidazole).

DETAILED DESCRIPTION

In a first embodiment, the present disclosure provides kinase inhibitorsof the Formula (I):

or a pharmaceutically acceptable salt thereof, wherein:

wherein

A is C or N;

B, D, and E are each independently selected from CR₀, N, O, and S;

R₀ is selected from H, —F, —Cl, —CH₃, —C₂-C₄ linear or branched alkyl,—C₂-C₄ alkenyl, —C₂-C₄ alkynyl, —CHR^(A)R^(B), —(CH₂)_(n)—Y,—CO(CH₂)_(n)Y, —(CH₂)_(n)— NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹,—NR^(A)R^(B), —OR², —CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂,—CF₃, —CF₂[18]F, —CF[18]F₂, and —C[18]F₃;

provided that R⁰ is not CF₃ when A, B, D, and E form a 1H-imidazolering;

Ar is a 6-membered substituted aryl, a 6-membered optionally substitutedheteroaryl, a 5 membered optionally substituted heteroaryl, or a5,6-fused bicyclic optionally substituted heteroaryl ring system;

R¹ is independently H, CH₃, —CH₂CH₃, or cyclopropyl;

R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or —(CH₂)_(n)—NR^(A)R^(B);

R^(A) and R^(B) are each independently H, CH₃, —CH₂CH₃, or cyclopropyl;R^(A) and R^(B) taken together form a 3-6 membered carbocyclic ringsystem or 5-7 membered saturated heterocyclic ring system;

Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹, —NR^(A)R^(B), —NR¹COR²,—S(O)_(m)R¹, —SO₂NR^(A)R^(B), —[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃,—[O—(CH₂)₂]_(n)—CH₂ ¹⁸F, —CH₂—F; —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂,—CF₃, —CF₂[18]F, —CF[18]F₂, or —C[18]F₃;

m is 0, 1, or 2;

n is 1, 2, or 3;

G is an appropriate group that interacts with the Gatekeeper region inthe ATP binding site of a specific kinase or profile of kinases ofinterest;

L is a linker group or ring system; and

(H) is an appropriate group that interacts with the Hinge- region in theATP binding site of a specific kinase or profile of kinases of interest.

In a second embodiment, the present disclosure provides kinaseinhibitors of the Formula (I), wherein R₀ is selected from H, —F, —Cl,—CH₃, —C₂-C₄ linear or branched alkyl, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CHR^(A)R^(B), —Y, —CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B),—[O—(CH₂)₂]_(n)Y, —(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹,—S(O)_(m)R¹, —NR^(A)R^(B), —OR², —CH₂—F, —CHF₂, and —CF₃; provided thatR⁰ is not CF₃ when A, B, D, and E form a 1H-imidazole ring; and Y is—CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹, —NR^(A)R^(B), —NR¹COR²,—S(O)_(m)R¹, —SO₂NR^(A)R^(B), —[O—(CH₂)₂]_(n)—CH₂F, —CH₂—F, —CHF₂, or—CF₃; and wherein the remaining variables are as described in the aboveembodiment.

In a third embodiment, the compounds of Formula (I) are represented bythe following structural formulae:

and pharmaceutically acceptable salts thereof, wherein X is —O—, —NR²—,or —S—; and wherein the remaining variables are as described in theabove embodiments.

In a fourth embodiment, Ar is of the structural formulae:

wherein:

Z is independently selected from —CH—, —CF—, or —N—;

X is selected from —O—, —S—, —NR²—;

R₃, R₄, and R₅ are each independently selected from H, —CH₃, —CH₂CH₃,cyclopropyl, —F, -[18]F, —Cl, —Br, —CH₂F, —CH₂[18]F, —CHF₂, —CF₃,—HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and —S(O)_(m)R¹; and wherein theremaining variables are as described in the above embodiments.Alternatively, R₃, R₄, and R₅ are each independently selected from H,—CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —Br, —CH₂F, —CHF₂, —CF₃, —HC═CHR¹,—CCR¹; —CN, —OCF₃, —NHR—OR¹, and —S(O)_(m)R¹.

In a fifth embodiment, G is of the structural formulae:

wherein:

R⁶, R⁷, and R⁸ are each independently selected from H, —CH₃, —CH₂CH₃,cyclopropyl, —CN; —OR¹, —(CH₂)_(n)OR¹, —NR^(A)R^(B),—(CH₂)_(n)NR^(A)R^(B), —S(O)_(m)R¹, —(CH₂)_(n)—S(O)_(m)R¹, —F, and —Cl;and wherein the remaining variables are as described in the aboveembodiments.

In a sixth embodiment, G is of structural formulae:

and wherein the remaining variables are as described in the aboveembodiments.

In a seventh embodiment, L is

i. a bond, —(CH₂)_(n)—, —(CH₂)_(n)—O—, -(cis)CH═CH—, -(trans)CH═CH—,—CC—, —OCH₂—, —OCH(CH₃)—, —CH(CH₃)O—, —S(O)_(m)—, —(CH₂)_(n)S(O)_(m)—,—S(O)_(m)(CH₂)_(n)—, —(CH₂)_(n)NH—, —NH—(CH₂)_(n)—, —NHCH(CH₃)—,—CH(CH₃)NH—, —NR₁C(O)—, —C(O)NR₁—, —XC(O)NH—, —NHC(O)X—, —(CH₂)_(n)—CO—,or —CO—(CH₂)_(n)—; or

ii. of the structural formulae

wherein R¹⁰ is hydrogen; and wherein the remaining variables are asdescribed in the above embodiments.

In an eighth embodiment, (H) is of the structural formulae:

wherein:

U is —H, F, Cl, —OR¹, or —NHR¹;

V and W are each independently selected from —H, —F, —Cl, —CF₃, —CONHR²,—X—R², —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹, X—(CH₂)_(m)CONR^(A)R^(B),—X—CH₂—(CH₂)OR¹, X—CH₂—(CH₂)NR^(A)R^(B), —X—CH₂—(CH₂)_(n)S(O)_(m)R¹,—X—(CH₂)_(n)S(O)_(m)NR^(A)R^(B), —O—(CH₂)₂NR^(A)R^(B),—O—(CH₂)₃NR^(A)R^(B), —O—(CH₂)_(n)CONR^(A)R^(B), —C₅-C₆heteroaryl,—COCH═CH—(CH₂)_(n)NR^(A)R^(B), and T; and

T is

and wherein the remaining variables are as described in the aboveembodiments.

In an ninth embodiment, (H) is alternatively of the structural formulae:

wherein the remaining variables are as described in the aboveembodiments.

In a tenth embodiment, compounds of the Formula (I) are represented bystructural Formulae (III) or (IV):

or pharmaceutically acceptable salt thereof, and wherein the remainingvariables are as described in the above embodiments.

In an eleventh embodiment, compounds described herein are represented bystructural Formulae (V) or (VI):

and pharmaceutically acceptable salts thereof, and wherein the remainingvariables are as described in the above embodiments.

In a twelfth embodiment, compounds described herein are represented bystructural Formulae (VII) to (XIV):

or a pharmaceutically acceptable salt, wherein R¹¹ is selected from H,—CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃, and —S—CH₃; and whereinthe remaining variables are as described in the above embodiments.

In a thirteenth embodiment, L is a bond, —O—, ═NH—. —S(O)_(m)—, —CC—,—O—CHR¹—, —NHCHR¹—, S(O)_(m)CHR¹—, —CHR¹O—, —CHR¹NH—, or —CHR¹S(O)_(m)—;and wherein the remaining variables are as described in the aboveembodiments.

In a fourteenth embodiment, L is of the structural formulae:

wherein R¹² is selected from H, —CH₃, —CH₂—CH₃, cyclopropyl,—S(O)_(m)R², —O—R², and NR^(A)R^(B); and wherein the remaining variablesare as described in the above embodiments.

In a fifteenth embodiment, (H) is of structural formulae:

wherein C₅-C₆ heteroaryls selected from

and wherein the remaining variables are as described in the aboveembodiments.

In a sixteenth embodiment, the compounds provided herein are of theformulae:

or a pharmaceutically acceptable salt thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃, —(CH₂)_(n)CH₂[18]F-[11]CH₃ or (CH₂)_(m)NR^(A)R^(B); andwherein the remaining variables are as described in the aboveembodiments. Alternatively, R₁₄ is —(CH₂)_(m)—CH₃, or—(CH₂)_(m)NR^(A)R^(B).

In a seventeenth embodiment, the compounds provided herein are of theformula:

and pharmaceutically acceptable salts thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃ or —(CH₂)_(m)NR^(A)R^(B); and wherein the remainingvariables are as described in the above embodiments.

In an eighteenth embodiment, the compounds provided herein are of theformula:

or a pharmaceutically acceptable salt thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃, —(CH₂)_(n)CH₂[18]F-[11]CH₃ or —(CH₂)_(m)NR^(A)R^(B); andwherein the remaining variables are as described in the aboveembodiments. Alternatively, R₁₄ is —(CH₂)_(m)—CH₃, or—(CH₂)_(m)NR^(A)R^(B).

In a nineteenth embodiment, the compounds provided herein are of theformula:

and pharmaceutically acceptable salts thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃, —(CH₂)_(n)CH₂[18]F-[11]CH₃ or —(CH₂)_(m)NR^(A)R^(B); andwherein the remaining variables are as described in the aboveembodiments. Alternatively, R₁₄ is —(CH₂)_(m)—CH₃, or—(CH₂)_(m)NR^(A)R^(B).

In a twentieth embodiment, the compounds provided herein are of theformula:

and pharmaceutically acceptable salts thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃, —(CH₂)_(n)CH₂[18]F-[11]CH₃ or —(CH₂)_(m)NR^(A)R^(B); andwherein the remaining variables are as described in the aboveembodiments. Alternatively, R₁₄ is —(CH₂)_(m) CH₃, or—(CH₂)_(m)NR^(A)R^(B).

In a twenty-first embodiment, the compounds provided herein are of theformula:

and pharmaceutically acceptable salts thereof, wherein R₁₄ is—(CH₂)_(m)—CH₃, —(CH₂)_(n)CH₂[18]F-[11]CH₃ or —(CH₂)_(m)NR^(A)R^(B); andwherein the remaining variables are as described in the aboveembodiments. Alternatively, R₁₄ is —(CH₂)_(m)—CH₃, or—(CH₂)_(m)NR^(A)R^(B).

In a twenty-second embodiment, the compounds provided herein are of thestructural formulae (XX-XXV):

and pharmaceutically acceptable salts thereof; wherein the variables areas described in the above embodiments. Alternatively, the compoundsprovided herein are of the structural formulae (XXVI-XXVIV):

and pharmaceutically acceptable salts thereof; wherein the variables areas described in the above embodiments.

In a twenty-third embodiment, L is —O—, —S—, —NH— or —C(O)NR¹; and (H)is of the structural formulae:

wherein the remaining variables are as described in the aboveembodiments.

In a twenty-fourth embodiment, L is —O— or —C(O)NR¹; and (H) is of thestructural formulae:

wherein the remaining variables are as described in the aboveembodiments.

In a twenty-fifth embodiment, (H) is of structural formulae:

In a twenty-sixth embodiment, the compounds described herein areselected from:

and pharmaceutically acceptable salts thereof.

In a twenty-seventh embodiment, the compounds described herein areselected from:

Specific kinase inhibitors and pharmaceutically acceptable salts thereofare provided in Examples below.

In an embodiment, the compounds described herein are useful forinhibiting kinases or a profile of kinases, such as, by administering toa subject an effective amount of one or more of the compounds describedherein. The kinases may be selected from e.g., Raf, PI3K, and P38, orcombinations thereof, and from type-III kinases members, including e.g.,CSF1R(FMS), PDGFR-α, PDGFR-β, KIT, and DDR kinases and/or C-Raf. Suchinhibition may be used for the treatment of inflammatory conditions andcancers. Additionally, the compounds described herein inhibit members ofthe TAM and/or MNK family of kinases and are useful in treating cancers,in particular resistant cancers, and for means of enhancingimmunotherapies.

Diseases and conditions treatable by the compounds described hereininclude, but are not limited to, chronic inflammatory conditions,neurodegenerative disorders, cancers, cardiovascular diseases,restenosis after percutaneous coronary intervention, venous bypass graftdisease, type-2 diabetes, infectious diseases, and neuropathic pain. Thecompounds described herein may also be used as positron emissiontomography tracers.

Specific examples of compounds of disclosed Formulae may exist invarious stereoisomeric or tautomeric forms. The invention encompassesall such forms, including active compounds in the form of essentiallypure enantiomers, racemic mixtures, and tautomers, including forms thosenot depicted structurally.

When any variable (e.g., aryl, heterocyclyl, R¹, R², etc.) occurs morethan once in a compound, its definition on each occurrence isindependent of any other occurrence.

The term “alkyl”, used alone or as part of a larger moiety such as“alkoxy”, “hydroxyalkyl”, “alkoxyalkyl”, “alkylamine”, “dialkyamine”,“alkoxycarbonyl” or “alkylaminocarbonyl”, means a saturated straight orbranched hydrocarbon radical having (unless otherwise specified) 1-10carbon atoms and includes, for example, methyl, ethyl, n-propyl,isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, n-hexyl,n-heptyl, n-octyl, n-nonyl, n-decyl and the like.

The term “cycloalkyl” means a saturated hydrocarbon ring having 3-8carbon atoms and includes, for example, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

The term “aryl” means means a 6-10 membered carbocyclic aromaticmonocyclic or polycyclic ring system, such as phenyl or naphthyl. Theterm “aryl” may be used interchangeably with the terms “aryl ring”“aromatic ring”, “aryl group” and “aromatic group”.

“Heteroaromatic group” or “heteroaryl” used alone or as part of a largermoiety as in “heteroaralkyl” or “heteroarylalkoxy”, means a 5-10membered monovalent monocyclic and polycylic aromatic group radicalcontaining 1 to 4 heteroatoms independently selected from N, O, and S.Heteroaryl groups include furyl, thienyl, thiophenyl, pyrrolyl,oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl,oxadiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridinyl-N-oxide,pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, indolyl, isoindolyl,benzo[b]furyl, benzo[b]thienyl, indazolyl, benzimidazolyl,benzthiazolyl, purinyl, 4H-quinolizinyl, quinolinyl, isoquinolinyl,quinazolinyl, benzothienyl, benzofuranyl, 2,3-dihydrobenzofuranyl,benzodioxolyl, benzimidazolyl, indazolyl, benzisoxazolyl, benzoxazolyl,benzothiazolyl, cinnolinyl, phthalzinyl, quinazolinyl, quinoxalinyl,1,8-naphthyridinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,3,4-oxadiazolyl,1,2,5-thiadiazolyl, 1,2,5-thiadiazolyl-1-oxide,1,2,5-thiadiazolyl-1,1-dioxide, 1,3,4-thiadiazolyl, 1,2,4-triazinyl,1,3,5-triazinyl, tetrazolyl, and pteridinyl. The terms “heteroaryl”,“heteroaromatic”, “heteroaryl ring”, “heteroaryl group” and“heteroaromatic group” are used interchangeably herein.

The term “heterocyclic group” or “heterocyclic ring” means a 4-, 5-, 6-and 7-membered saturated or partially unsaturated heterocyclic ringcontaining 1 to 4 heteroatoms independently selected from N, O, and S,and include pyrrolidine, piperidine, tetrahydrofuran, tetrahydropyran,tetrahydrothiophene, tetrahydrothiopyran, isoxazolidine, 1,3-dioxolane,1,3-dithiolane, 1,3-dioxane, 1,4-dioxane, 1,3-dithiane, 1,4-dithiane,morpholine, thiomorpholine, thiomorpholine 1,1-dioxide,tetrahydro-2H-1,2-thiazine 1,1-dioxide, and isothiazolidine 1,1-dioxide.The terms “heterocyclyl”, “heterocycle”, “heterocyclic group” and“heterocyclic ring” are used interchangeably herein.

Suitable substituents for a aryl, heteroaryl and heterocyclic group arethose which do not significantly reduce the ability of the compound toinhibit the activity of kinases. Unless otherwise specified, suitablesubstituents for an aryl, heteroaryl and heterocyclic group includehalo, OR¹¹, S(O)_(p)R¹¹, CN, NO₂, C(O)R¹¹, C(S)R¹¹, CO₂R¹¹, CHO,(C₁-C₆)alkyl, halo(C₁-C₆)alkyl, halo(C₁-C₆)alkoxy, NR¹¹R¹², CONR¹¹R¹²,OC(O)NR¹¹R¹², NR¹¹C(O)NR¹¹R¹², CSNR¹¹R¹², OC(S)NR¹¹R¹², NR¹¹C(S)NR¹¹R¹²,SO₂NR¹¹R¹², NR¹¹SO₂NR¹¹R¹², NR¹¹C(O)R¹², OC(O)R¹², NR¹¹C(S)R¹²,NR¹¹C(O)OR¹³, NR¹¹C(S)OR³, or NR¹¹SO₂R³; or a (C₁-C₆)alkyl substitutedwith halo, OR¹¹, S(O)_(p)R¹¹, CN, NO₂, C(O)R¹¹, C(S)R¹¹, CO₂R¹¹, CHO,(C₁-C₆)alkyl, halo(C₁-C₆)alkyl, NR¹¹R¹², CONR¹¹R¹², OC(O)NR¹¹R¹²,NR¹¹C(O)NR¹¹R¹², CSNR¹¹R¹², OC(S)NR¹¹R¹², NR¹¹C(S)NR¹¹R¹², SO₂NR¹¹R¹²,NR¹¹SO₂NR¹¹R¹², NR¹¹C(O)R¹², OC(O)R¹², NR¹¹C(S)R¹², NR¹¹C(O)OR³,NR¹¹C(S)OR³, or NR¹¹SO₂R³, wherein R¹¹⁻¹³ are each independentlyC₁-C₆alkyl; C₁-C₆haloalkyl, or C₁-C₆hydroxyalkyl. Preferred substituentsan alkyl, aryl, heteroaryl and heterocyclyl include, unless otherwisespecified, halogen, (C₁-C₆)alkyl, hydroxy, (C₁-C₆)alkoxy,(C₁-C₆)alkylamino, di(C₁-C₆)alkylamino, NO₂, CN, CONH₂, (C₁-C₆)haloakylor (C₁-C₆)haloalkoxy.

An appropriate group that interacts with the Gatekeeper region in theATP binding side includes either an optionally substituted planarmonocyclic or bicyclic aromatic groups or rarely cyclic aliphaticgroups. These lipophilic groups interact with the lipophilic pocketadjacent to the ATP purine binding site of a kinase. Many small-moleculekinase inhibitors have exploited a conserved threonine residue withinthe ATP binding site for binding specificity. This threonine controlsaccess of the inhibitors to a hydrophobic pocket deep in the active sitethat is not contacted by ATP, hence leading to its designation as a‘gatekeeper’ residue. Substitution of the gatekeeper threonine residuewith bulky side chains is a common mechanism of resistance topharmacological ATP-competitive kinase inhibitors. Representativegatekeeper interacting groups have been defined earlier.

An appropriate group that interacts with the Hinge-region in the ATPbinding site includes optionally substituted planar monocyclic orpolycyclic aromatic ring systems with a network of hydrogen-bondacceptors and donors that are complimentary to the conserved hingeregion that interact with the purine ring of ATP. RepresentativeHinge-region interacting groups are illustrated below.

The compounds described herein may be present in the form ofpharmaceutically acceptable salts. For use in medicines, the salts ofthe compounds of the invention refer to non-toxic “pharmaceuticallyacceptable salts.” Pharmaceutically acceptable salt forms includepharmaceutically acceptable acidic/anionic or basic/cationic salts.

Pharmaceutically acceptable acidic/anionic salts include, the acetate,benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide, calciumedetate, camsylate, carbonate, chloride, citrate, dihydrochloride,edetate, edisylate, estolate, esylate, fumarate, glyceptate, gluconate,glutamate, glycollylarsanilate, hexylresorcinate, hydrobromide,hydrochloride, hydroxynaphthoate, iodide, isethionate, lactate,lactobionate, malate, maleate, mandelate, mesylate, methylsulfate,mucate, napsylate, nitrate, pamoate, pantothenate, phosphate/diphospate,polygalacturonate, salicylate, stearate, subacetate, succinate, sulfate,tannate, tartrate, teoclate, tosylate, and triethiodide salts.

The compounds described herein may also include pharmaceuticallyacceptable anionic salt forms, wherein the anionic salts include theacetate, benzenesulfonate, benzoate, bicarbonate, bitartrate, bromide,calcium edetate, camsylate, carbonate, chloride, citrate,dihydrochloride, edetate, edisylate, estolate, esylate, fumarate,glyceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate,hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isethionate,lactate, lactobionate, malate, maleate, mandelate, mesylate,methylsulfate, mucate, napsylate, nitrate, pamoate, pantothenate,phosphate/diphospate, polygalacturonate, salicylate, stearate,subacetate, succinate, sulfate, tannate, tartrate, teoclate, tosylate,and triethiodide salts.

Salts of the disclosed kinase inhibitors containing an acidic functionalgroup can be prepared by reacting with a suitable base. Such apharmaceutically acceptable salt may be made with a base which affords apharmaceutically acceptable cation, which includes alkali metal salts(especially sodium and potassium), alkaline earth metal salts(especially calcium and magnesium), aluminum salts and ammonium salts,as well as salts made from physiologically acceptable organic bases suchas trimethylamine, triethylamine, morpholine, pyridine, piperidine,picoline, dicyclohexylamine, N,N′-dibenzylethylenediamine,2-hydroxyethylamine, bis-(2-hydroxyethyl)amine,tri-(2-hydroxyethyl)amine, procaine, dibenzylpiperidine,dehydroabietylamine, N,N′-bisdehydroabietylamine, glucamine,N-methylglucamine, collidine, quinine, quinoline, and basic amino acidssuch as lysine and arginine.

When a disclosed compound or its pharmaceutically acceptable salt isnamed or depicted by structure, it is to be understood that solvates orhydrates of the compound or its pharmaceutically acceptable salts areincluded as well as anhydrous forms of the compound and forms withoutsolvent. “Solvates” refer to crystalline forms wherein solvent moleculesare incorporated into the crystal lattice during crystallization.Solvate may include water or nonaqueous solvents such as ethanol,isopropanol, DMSO, acetic acid, ethanolamine, and EtOAc. Solvates,wherein water is the solvent molecule incorporated into the crystallattice, are typically referred to as “hydrates”. Hydrates includestoichiometric hydrates as well as compositions containing variableamounts of water.

When a disclosed compound or its pharmaceutically acceptable salt isnamed or depicted by structure, it is to be understood that thecompound, including solvates thereof, may exist in crystalline forms,non-crystalline forms or a mixture thereof. The compound or itspharmaceutically acceptable salts or solvates may also exhibitpolymorphism (i.e. the capacity to occur in different crystallineforms). These different crystalline forms are typically known as“polymorphs.” It is to be understood that when named or depicted bystructure, the disclosed compound and its pharmaceutically acceptablesalts, solvates or hydrates also include all polymorphs thereof.Polymorphs have the same chemical composition but differ in packing,geometrical arrangement, and other descriptive properties of thecrystalline solid state. Polymorphs, therefore, may have differentphysical properties such as shape, density, hardness, deformability,stability, and dissolution properties. Polymorphs typically exhibitdifferent melting points, IR spectra, and X-ray powder diffractionpatterns, which may be used for identification. One of ordinary skill inthe art will appreciate that different polymorphs may be produced, forexample, by changing or adjusting the conditions used in solidifying thecompound. For example, changes in temperature, pressure, or solvent mayresult in different polymorphs. In addition, one polymorph mayspontaneously convert to another polymorph under certain conditions.

The present disclosure also includes various isomers and mixturesthereof. “Isomer” refers to compounds that have the same composition andmolecular weight but differ in physical and/or chemical properties. Thestructural difference may be in constitution (geometric isomers) or inthe ability to rotate the plane of polarized light (stereoisomers).

Certain disclosed kinase inhibitors may exist in various stereoisomericforms. Stereoisomers are compounds that differ only in their spatialarrangement. Enantiomers are pairs of stereoisomers whose mirror imagesare not superimposable, most commonly because they contain anasymmetrically substituted carbon atom that acts as a chiral center.“Enantiomer” means one of a pair of molecules that are mirror images ofeach other and are not superimposable. Diastereomers are stereoisomersthat are not related as mirror images, most commonly because theycontain two or more asymmetrically substituted carbon atoms. The symbol“*” in a structural formula represents the presence of a chiral carboncenter. “R” and “S” represent the configuration of substituents aroundone or more chiral carbon atoms. Thus, “R*” and “S*” denote the relativeconfigurations of substituents around one or more chiral carbon atoms.When a chiral center is not defined as R or S, a mixture of bothconfigurations is present.

“Racemate” or “racemic mixture” means a compound of equimolar quantitiesof two enantiomers, wherein such mixtures exhibit no optical activity;i.e., they do not rotate the plane of polarized light.

“Geometric isomer” means isomers that differ in the orientation ofsubstituent atoms in relationship to a carbon-carbon double bond, to acycloalkyl ring, or to a bridged bicyclic system. Atoms (other than H)on each side of a carbon-carbon double bond may be in an E (substituentsare on opposite sides of the carbon-carbon double bond) or Z(substituents are oriented on the same side) configuration.

The compounds of the invention may be prepared as individual isomers byeither isomer-specific synthesis or resolved from an isomeric mixture.Conventional resolution techniques include forming the salt of a freebase of each isomer of an isomeric pair using an optically active acid(followed by fractional crystallization and regeneration of the freebase), forming the salt of the acid form of each isomer of an isomericpair using an optically active amine (followed by fractionalcrystallization and regeneration of the free acid), forming an ester oramide of each of the isomers of an isomeric pair using an optically pureacid, amine or alcohol (followed by chromatographic separation andremoval of the chiral auxiliary), or resolving an isomeric mixture ofeither a starting material or a final product using various well knownchromatographic methods.

When a disclosed compound is named or depicted by structure withoutindicating the stereochemistry, and the compound has at least one chiralcenter, it is to be understood that the name or structure encompassesone enantiomer of inhibitor free from the corresponding optical isomer,a racemic mixture of the inhibitor and mixtures enriched in oneenantiomer relative to its corresponding optical isomer.

When the stereochemistry of a disclosed compound is named or depicted bystructure, the named or depicted stereoisomer is at least 60%, 70%, 80%,90%, 95%, 98%, 99% or 99.9% by weight pure relative to the otherstereoisomers. When a single enantiomer is named or depicted bystructure, the depicted or named enantiomer is at least 60%, 70%, 80%,90%, 95%, 98%, 99% or 99.9% by weight optically pure. Percent opticalpurity by weight is the ratio of the weight of the enantiomer over theweight of the enantiomer plus the weight of its optical isomer. When asingle geometric isomer, e.g., a geometric isomer with a double bond, isdepicted by name or structure and the stereochemistry about the doubleis indicated, the compound is considered to be at least 60%, 70%, 80%,90%, 95%, 98%, 99% or 99.9% steroechemically pure by weight. Percentstereochemically purity by weight is the ratio of the weight of thegeometric isomer over the weight of the both geometric isomers. Forexample, 99% stereochemically pure means that at least 99% by weight ofthe compound is the indicated stereoisomer.

A pharmaceutical composition of the compounds disclosed herein may,alternatively or in addition to a compound of the above formulae,comprise a pharmaceutically acceptable salt of a compound of the aboveformulae, or a prodrug or pharmaceutically active metabolite of such acompound or salt and one or more pharmaceutically acceptable carrierstherefor.

“Effective amount” means that amount of active compound agent thatelicits the desired biological response in a subject. Such responseincludes alleviation of the symptoms of the disease or disorder beingtreated.

“Inhibiting kinase activity” means to decrease the production of anphosphor-peptide by a particular kinase that can be separated byanalytical techniques. For example, a comprehensive platform of kinase,protease, phosphatase and epigenetic assays that utilizes cutting-edgemicrofluidics and informatics tools to provide accurate and rapidresults. Profiling data in this application were determined using thesetechniques at Nanosyn Inc. IC50 data were provided by Reaction BiologyInc using a P33 radiolabeled kinase assay.

“Pharmaceutically acceptable carrier” means compounds and compositionsthat are of sufficient purity and quality for use in the formulation ofa composition of the present disclosure and that, when appropriatelyadministered to an animal or human, do not produce an adverse reaction.

“Treatment” or “treating”, as used herein, includes therapeutictreatment. “Therapeutic treatment” includes partially or totallyinhibiting, delaying, or reducing the severity of the disease ordisorder related to kinases. “Prophylactic treatment” encompassesadministration of a compound of the invention to a subject susceptibleto a disease or disorder related to the activity or expression ofkinases in an effort to reduce the likelihood of a subject developingthe disease or disorder, or slowing or preventing progression of thedisease. Prophylactic treatment includes suppression (partially orcompletely) of the disease or disorder, and further includes reducingthe severity of the disease or disorder, if onset occurs. Prophylactictreatment is particularly advantageous for administration to mammals atrisk for developing a disease or disorder related to kinase activity.

The compounds disclosed herein can be prepared and administered in awide variety of oral and parenteral dosage forms. Thus, the compounds ofthe present disclosure can be administered by injection, that is,intravenously, intramuscularly, intracutaneously, subcutaneously,intraduodenally, or intraperitoneally. Additionally, the compounds ofthe present disclosure can be administered intranasally ortransdermally.

For preparing pharmaceutical compositions from the compounds of thepresent disclosure, pharmaceutically acceptable carriers can either besolid or liquid. Solid form preparations include powders, tablets,pills, capsules, cachets, suppositories, and dispersable granules. Asolid carrier can be one or more substances which may also act asdiluents, flavoring agents, solubilizers, lubricants, suspending agents,binders, preservatives, tablet disintegrating agents, or anencapsulating material. In powders, the carrier is a finely dividedsolid which is in a mixture with the finely divided active ingredient.

In tablets, the active ingredient is mixed with the carrier having thenecessary binding properties in suitable proportions and compacted inthe shape and size desired.

The powders and tablets preferably contain from about one to aboutseventy percent of the active ingredient. Suitable carriers aremagnesium carbonate, magnesium stearate, talc, sugar, lactose, pectin,dextrin, starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcelluose, a low melting wax, cocoa butter, and the like.Tablets, powders, cachets, lozenges, fast-melt strips, capsules andpills can be used as solid dosage forms containing the active ingredientsuitable for oral administration.

For preparing suppositories, a low melting wax, such as a mixture offatty acid glycerides or cocoa butter, is first melted and the activeingredient is dispersed homogeneously therein, as by stirring. Themolten homogeneous mixture is then poured into convenient sized molds,allowed to cool, and thereby to solidify.

Liquid form preparations include solutions, suspensions, retentionenemas, and emulsions, for example, water or water propylene glycolsolutions. For parenteral injection, liquid preparations can beformulated in solution in aqueous polyethylene glycol solution.

Aqueous solutions suitable for oral administration can be prepared bydissolving the active ingredient in water and adding suitable colorants,flavors, stabilizing, and thickening agents as desired. Aqueoussuspensions for oral administration can be prepared by dispersing thefinely divided active ingredient in water with viscous material, such asnatural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, and other well-known suspending agents.

The pharmaceutical composition is preferably in unit dosage form. Insuch form, the composition is subdivided into unit doses containingappropriate quantities of the active ingredient. The unit dosage formcan be a packaged preparation, the package containing discretequantities of, for example, tablets, powders, and capsules in vials orampules. Also, the unit dosage form can be a tablet, cachet, capsule, orlozenge itself, or it can be the appropriate amount of any of these inpackaged form.

The quantity of active ingredient in a unit dose preparation may bevaried or adjusted from about 0.1 mg to about 1000.0 mg, preferably fromabout 0.1 mg to about 100 mg. The dosages, however, may be varieddepending upon the requirements of the patient, the severity of thecondition being treated, and the compound being employed. Determinationof the proper dosage for a particular situation is within the skill inthe art. Also, the pharmaceutical composition may contain, if desired,other compatible therapeutic agents.

The compounds provided herein are useful for ameliorating or treatingdisorders or diseases in which inhibition of a kinase or profile ofkinases has utility for the treatment of certain cancers for whichactivating mutations in, over expression of, or abberant activity ofthese kinase targets contribute to the progression of the disorder ordisease.

The compounds provided herein are also useful as multi-targeted kinaseinhibitors of the Protein Tyrosine Kinases (PTK), Receptor TyrosineKinases, (RTK), Serine/Theonine Kinases. And alipid kinases such as thePI3K kinase family. As such, these inhibirors will have utility for thetreatment of cancers and/or inflammatory conditions sensitive to kinaseinhibitors. In particular, the ability to inhibit MAP-kinases such asp38 and Raf kinases in combination with PI3K kinases is novel and offersadvantages for unique therapies not previously available.

Compounds described herein, that inhibit a kinase or profile of relevantkinases will have utility for the treatment of chronic inflammatoryconditions such as rheumatoid arthritis; neurodegenerative disorderssuch as Alzheimer's disease; cardiovascular diseases such asatherosclerosis, restenosis after percutaneous coronary intervention,and venous bypass graft disease; type-2 diabetes; certain infectiousdiseases such as parasitic infections; and neuropathic pain, for whichover expression or aberrant activity of specific kinase targetscontribute to progression of the disease.

Compounds described herein, also have utility as positron emissiontomography tracers, such as those in FIG. 12 and FIG. 13. These agentswill enable diagnostic PET imaging studies that will have utilities forthe identification, characterization, and monitoring of diseases inwhich the mutation or aberrant activity of diagnostically relevantkinase targets as well as selection of appropriate patient populationsfor clinical studies.

Compounds described herein which have utility, when attached to anappropriate vehicle through the free amine group, as affinity probes forthe identification of kinases that can adopt the DFG-out conformation,such as those illustrated in FIG. 11. These agents will enable theidentification and kinetic study of new kinases for which Type-IIinhibitors have thus far not been reported.

Compounds, methods, and utilities described herein may be used as aplatform technology for the patient specific study of kinase inhibitorsand their efficacy in the clinic. In particular, this disclosuredescribes how the chemistry methods for preparation of multi-targetedkinase inhibitors can be easily adapted to the preparation ofradiolabeled tracers for PET imaging. The integrated use of PET imagingprobes with very similar properties and inhibition profiles as thetherapeutic agent of interest will offer significant and uniqueadvantages for selection of most appropriate patient populations forclinical studies. Targeted monitoring of kinases using PET imagingrepresents a significant step toward the realization of personalizedmedicine

The term “mammal” is preferably a human, but can also be an animal inneed of veterinary treatment, e.g., companion animals (e.g., dogs, cats,and the like), farm animals (e.g., cows, sheep, pigs, horses, and thelike) and laboratory animals (e.g., rats, mice, guinea pigs, and thelike).

The following abbreviations have the indicated meanings:

Abbreviation Meaning Boc tert-butoxy carbonyl or t-butoxy carbonyl(Boc)₂O di-tert-butyl dicarbonate BOP(Benzotriazole-1-yl-oxy-tris-(dimethylamino)- phosphoniumhexafluorophosphate) Cbz Benzyloxycarbonyl CbzCl Benzyl chloroformateCS₂ Carbon disulfide DAST diethylaminosulfur trifluoride DBU1,8-diazabicyclo[5.4.0]undec-7-ene DCC N,N′-dicyclohexylcarbodiimide DCMdichloromethane DCU N,N′-dicyclohexylurea DIAD diisopropylazodicarboxylate DIEA N,N-diisopropylethylamine DMAP4-(dimethylamino)pyridine DMF N,N-dimethylformamide DMPU1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone 2,4-DNP2,4-dinitrophenylhydrazine DPTBS Diphenyl-t-butylsilyl EDC, EDC•HCl,1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide EDCI hydrochloride EquivEquivalents Fmoc 1-[[(9H-fluoren-9-ylmethoxy)carbonyl]oxy]- Fmoc-OSu1-[[(9H-fluoren-9-ylmethoxy)carbonyl]oxy]-2,5- pyrrolidinedione h, hrhour(s) HOAc acetic acid HOBt 1-hydroxybenzotriazole HATU2-(7-Aza-1H-benzotriazole-1-yl)-1,1,3,3- tetramethyluroniumhexafluorophosphate HBTU2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphateI₂ iodine KHMDS potassium hexamethyldisilazane LAH or LiAlH₄ lithiumaluminum hydride LC-MS liquid chromatography-mass spectroscopy LHMDSlithium hexamethyldisilazane Lawesson’s2,4-bis(4-methoxyphenyl)-1,3,2,4-dithiadiphosphetane- Reagent2,4-disulfide MCPBA: Meta-chloroperoxybenzoic acid Me Methyl MsClmethanesulfonyl chloride Min Minute MS mass spectrum NaH sodium hydrideNaHCO₃ sodium bicarbonate NaN₃ sodium azide NaOH sodium hydroxide NaNO₂sodium nitrite Na₂SO₄ sodium sulfate NH₃ ammonia NMM N-methylmorpholineNMP N-methylpyrrolidinone Oxone potassium monopersulfate Pd₂(dba)₃tris(dibenzylideneacetone)dipalladium(0) PE petroleum ether PhI +(OCOCF₃)₂ bis(trifluoroacetoxy)iodo]benzene PPh₃ triphenyl phospheneQuant quantitative yield Satd Saturated SOCl₂ thionyl chloride SFCsupercritical fluid chromatography SPA scintillation proximity assay SPEsolid phase extraction TBAF tetrabutylammonium fluoride TBSt-butyldimethylsilyl TBDPS t-butyldiphenylsilyl TBSClt-butyldimethylsilyl chloride TBDPSCl t-butyldiphenylsilyl chloride TEAtriethylamine or Et₃N TEMPO 2,2,6,6-tetramethyl-1-piperidinyloxy freeradical Teoc 1-[2-(trimethylsilyl)ethoxycarbonyloxy]- Teoc-OSu1-[2-(trimethylsilyl)ethoxycarbonyloxy]pyrrolidin- 2,5-dione TFAtrifluoroacetic acid TFAA trifluoroacetic anhydride Tlc, TLC thin layerchromatography THF tetrahydrofuran TMS Trimethylsilyl TMSClchlorotrimethylsilane or trimethylsilyl chloride t_(R) retention timeTsOH p-toluenesulfonic acid

EXEMPLIFICATION General Description of Synthetic Methods

Compounds disclosed herein can be prepared by several processes. In thediscussion below, the variables have the meanings indicated above unlessotherwise noted. In cases where the synthetic intermediates and finalproducts described below contain potentially reactive functional groups,for example amino, hydroxyl, thiol and carboxylic acid groups, that mayinterfere with the desired reaction, it may be advantageous to employprotected forms of the intermediate. Methods for the selection,introduction and subsequent removal of protecting groups are well knownto those skilled in the art. (T. W. Greene and P. G. M. Wuts “ProtectiveGroups in Organic Synthesis” John Wiley & Sons, Inc., New York 1999).Such protecting group manipulations are assumed in the discussion belowand not described explicitly. Generally, reagents in the reactionschemes are used in equimolar amounts; however, in certain cases it maybe desirable to use an excess of one reagent to drive a reaction tocompletion. This is especially the case when the excess reagent can bereadily removed by evaporation or extraction. Bases employed toneutralize HCl in reaction mixtures are generally used in slight tosubstantial excess (1.05-5 equivalents).

Preparation of Ortho-Aryl-5-Membered-Heteroaryl Carboxylic AcidScaffolds

The following paragraphs outline similar and alternative scaffolds andtheir preparation in order to define the scope of this invention.

Kinase inhibitors and imaging agent precursors described herein may beprepared by coupling of an appropriate substituted ortho-aryl-5-memberedheterocyclic carboxylic acid scaffold with an appropriate aminecontaining intact Hinge-Linker-Gatekeeper motif (HGM), FIG. 14. Standardcoupling conditions such as EDC/HOAT/DIEA or HATU/DIEA can be used toaffect this transformation (Route-1). Other methods of construction mayinvolve coupling with a partially elaborated HGM which can then befurther modified to give the final product. For example, in Route-2 anappropriately protected gatekeeper amine may be coupled to the scaffoldacid. If needed, deprotection of the resulting amide intermediate andcoupling of a optionally protected Hinge-Linker group would give theassembled product. Alternatively, the scaffold acid could be coupled toan appropriately protected Linker-Gatekeeper amine which, afterdeprotection if needed, would be coupled to an appropriately protectedHinge interacting group P—(H). Depending on the protection state ofthese assembled product, a final deprotection step may be required.

General methods for the preparation of a variety of scaffolds areillustrated in Schemes 1-12.

2-Substituted 5-aryl-imidazole-4-carboxylic acid scaffolds can beprepared as illustrated in Scheme 2. In Route-1, a racemic phenylglycine 1 is first converted to a variety of amides, ureas, orcarbamates derivatives 2. Cyclo-dehydration with trifluoro aceticanhydride (TFAA) provides oxazolidinone 3 which can be isolated andpurified or utilized directly in the same flask after removal ofvolatiles in vacuo. Reaction of 3 with benzyl cyanoformate and tributylphosphine in toluene in the same flask smoothly provides 2-substitutedimidazoles 4 via [3+2]cycloaddition reaction accompanied bydecarboxylative aromatization (Hagiwara, K. et. al., WO1995004724).

Route-2 represents a special case of this process in which aphenylglycine derivative 1 is first treated with excess oftrifluoroacetic anhydride. Formation of the intermediatetrifluoroacetamide 6 is accompanied by in situ cyclo-dehydration toprovide oxazolidinone 7 which can be isolated or utilized directly inthe same flask after removal of volatiles in vacuo. Reaction of 7 asdescribed in Route-1 provides 2-trifluoromethyl imidazole 8 in athree-step one-pot procedure. Hydrogenolysis or hydrolysis of the benzylesters then results in the desired imidazole carboxylic acid scaffold 5and 9 respectively. Some of the previously reported 2-trifluoromethylimidazoles scaffolds S-1 to S-8 prepared by this method are listed inTable 11 (Deitrich J., Bioorg. Med. Chem. (2010), 18(1): 292-304.).

TABLE 11 Previously Prepared 2-CF₃-Imidazole Scaffolds. (Dietrich 2010)Scaffold Scaffold Available Structure ID # Comment Source or Reference.Order # Precursor ID #

S-1 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-2 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-3 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-4 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-5 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-6 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-7 Dietrich, J.; Gokhale, V. Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

S-8 Dietrich, J.; Gokhale, V.; Wang. X.; Hurley, L. H. and Flynn, G. A.Bioorg, Med. Chem., 2010, 18, 292-304.

Alternatively, a more traditional general approach to2,4,5-trisubstituted imidazoles can be employed, Scheme 3. In Route-1,an aroylacetic ester 10 is halogenated (Lee L. F., J. Hetrocyclic Chem.(1985), 22(6): 1621-1630) followed by reaction of the 2-halointermediate 11 with an amidine derivative to provide a variety ofimidazole esters of general structure 12 that can then be hydrolyzed toprovide 2-substituted imidazole carboxylic acid scaffolds 13.Alternatively, the aroylacetic ester 10 can be treated with sodiumnitrite in acetic acid to provide an intermediate oxime 14 that can bereduced to 2-amino keto ester 15. Reaction of 15 with an imidate salt 16provides an efficient complimentary entrance to imidazole esters 12.

A variety of imidazolidin-2-one acid scaffolds are commerciallyavailable, Table 12, or can be easily accessed synthetically, Scheme 4.For example, reaction of chloro-ketoester 11 with urea (Route-1),provides a facile entrance to imidazolidin-2-oness 16 and reaction ofamine 15 with an isocyanate affords the N-substituted derivative. 18(Route-2).

TABLE 12 Commercially available imidazole-2-one scaffolds of interest.Scaffold Scaffold Available Structure ID # Comment Source or Reference.Order # Precursor ID #

S-9 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www.bestpharmatech.com BP21116

S-10 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www.bestpharmatech.com BP21165

S-11 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www.bestpharmatech.com BP21194

S-12 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www.bestpharmatech.com BP19184

S-13 Prepared from Precursor P-13 by LiOH ester hydrolysis

P-13

Alternative chemistry for the preparation and further conversion ofmercapto-imidazoles is outlined in Scheme 5. Reaction of2-amino-ketoester 15 with potassium isocyanate is known to be a verygeneral synthetic approach to 2-mercapto-imidazoles 19, Route-1. (Maeda,1984). Alternatively, reaction of 15 with and isothiocyanate yields theN-substituted derivative 20. Alkylation of 2-mercaptoimidaozles 19 andsubsequent partial or complete oxidation proceeds smoothly to yield the2-alkylthio-, 2-alkylsulfinyl-, and 2-alkylsulfonyl-imidazole esters 21,22, and 23 as intermediates to their corresponding acid scaffolds.,

TABLE 13 2-Mercapto-imidazole scaffolds prepared in this application.Scaffold Scaffold Order Available Structure ID # Comment Source orReference. # Precursor ID #

S-14 Maeda S. et.al. Synthesis of 2- mercapto-4-substituted imidazolederivatives with antiinflammatory activities. Chem. Pharm. Bull., 198432(7), 2536-43

P-14

S-15 Prepared from P-14 by alkylation with MeI then LiOH hydrolysis. Seeexperimental for S-15

S-16 Prepared from S-15 by treatment with MCPBA. See experimental forS-16

S-17 Prepared from S-15 by treatment with Oxone. See experimental forS-17

The 2-mercapto-imidazole scaffolds in Table 13 have been prepared, seeexperimental section. Reaction of 2-mercaptoimidazole P-14 with methyliodide followed by ester hydrolysis provides the 2-methylmercaptoscaffold S-15. Whereas partial oxidation of S-15 with MCPBA provides theenantiomeric sulfoxides S-16, oxidation P-15 with oxone and subsequentester hydrolysis provides the corresponding sulfone S-17.

A variety of 2-Substituted 5-aryl-thiazole-4-carboxylic acid scaffolds30-33 can be prepared by reacting a 3-chloro-3-arylpyruvate ester 25with an appropriate thioamide or thiourea (PCT Int. Appl., 2009 016560.5 Feb., 2009) to provide the 2-substituted thiazoles 26, or2-amino-thiazole if R is NH₂. Route-1 in Scheme 6. Alternatively,reaction of a 3-chloro-3-arylpyruvate ester 2 with the condensationproduct between carbon disulfide and ammonia is reported to provide thecorresponding thione intermediate 27, Route-2. Alkylation of thioneester 27 provides 2-alkylthio-5-aryl-thiazole-4-carboxylic esters 28that can be optionally oxidized to provide thiazole scaffolds witheither the sulfoxide and sulfone oxidation state 29. In an alternateapproach to thione 27, a 2-amino-aroylacetic ester 15 is condensed withcarbon disulfide and cyclo-dehydrated under acidic conditions to give27, Route-3. Because a diverse set of intermediate3-chloro-3-arylpyruvate esters 25 can easily be prepared from readilyavailable aryl aldehydes 24 via a modified Knoevenagel condensation,this general approach to 2-substituted-5-aryl-4-thiazole carboxylic acidscaffolds is quite general.

A variety of 2-mercapto-thiazole acid scaffolds S-18 to S-24 and estersP-21 and P-25 are commercially available, Table 14. Alkylation of P-25with methyl iodide and ester hydrolysis provides scaffold S-25.Oxidation of S-25 with 1 or 2 equivalents of m-chlorobenzoic acidprovides the respective sulfoxide and sulfone scaffolds S-26 and S-27,albeit in impure form, for S-25, S-26, and S-27.

TABLE 14 Available 2-substituted thiazole scaffolds S-18 to S-27.Scaffold Availabe Structure Scaffold ID # Comment Source or Reference.Order # Precursor ID #

S-18 ZereneX Molecular Limited 12 Manningham Road Greater Manchester,BL3 5QS United Kingdom www.zerenex-molecular.com ZXA008039

S-19 ZereneX Molecular Limited 12 Manningham Road Greater Manchester,BL3 5QS United Kingdom www.zerenex-molecular.com ZXA008069

S-20 Ryan Scientific, Inc. P O BOx 703 Mt. Pleasant, SC, 29465 USAEmail: sales@ryansci.com Web: http://www.ryansci.com CA00660

S-21 Preparation of ester precursor P-26: see PCT Int. Appl.,2009104155, 27 Aug. 2009

P-21

S-22 Otava Building Blocks Otava 55 Ellerslie Avenue, Suite 524 Toronto,ON, M2N 1X9 Canada http://www.otavachemicals.com 1805700

S-23 See experimental for S-23

S-24 ZereneX Molecular Limited 12 Manningham Road Greater Manchester,BL3 5QS United Kingdom www. zerenex-molecular.com ZXA008054

S-25 Prepard from Precursor P-25 with Mel and LiOH ester hydrolysisKingsh Chemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing, 210036People's Republic of China http://www.kingschem.com 42487

P-25

S-26 Prepared from S-25 by treatment with 1 eq. MCPBA See experimentalfor S-26

S-27 Prepared from S-25 by treatment with 2 eq. MCPBA See experimentalfor S-27

Analogous chemistry, using a 2-halo-aroylacetic ester 11 as the commonstarting material, can be applied to prepare 2-Substituted4-aryl-thiazole-5-carboxylic acid scaffolds (Zawadzka, Acta PoloniaePharmaceutica (1984), 41(6), 633-640), Scheme 7. Reaction of 11 with athiourea, Route-1 (Narender M, Synthesis, (2007) 3469-3472); athioamide, Route-2; or the adduct of carbon disulfide with ammonia,Route-3; affords the appropriate 2-substitute thiazole esters 34, 35,and 36, respectively. The 2-amino group of thiazole ester 3 can befurther functionalized on nitrogen or converted to 2-halo derivative 37through diazatization the action with the appropriate copper halidesalt. A variety of 2-substituted thiazoles 38 can then be prepared bynucleophilic aromatic substitution or transition metal mediated couplingreactions. (Lee 1986). Thiazolidinethione 36 can be alkylated andoptionally oxidized to provide 2-alkylmercapto-, 2-alkylsulfinyl-, or2-alkylsulfonyl-thiazole 5-carboxylic esters 39. Route-3. Using thesemethods, a variety of 2-substituted 4-aryl-thiazole-5-carboxylic acidscaffolds 40-45 can be prepared. This approach has been expanded toallow the preparation of alternative 2-substituted thiazoles. Thiazole2-carboxylic acid derivatives such as 46 may be prepared in an analogousmanner, Route-4 (Lilienkampf A., J. Med. Chem. (2009) 52: 2109-2118).

A variety of functionalized 2-aminothiazole scaffolds can be preparedfrom commercially available 2-aminothiazole intermediates such as P-29,Table 15.

TABLE 15 Sources for 2-aminothiazole scaffolds. Scaff- Scaffold oldAvailable Structure ID # Comment Source or Reference. Order # PrecursorID #

S-28 Atlantic Research Chemicals Ltd. Unit A, Stratton View BusinessPark Stratton Bude, Cornwall EX23 9NR United Kingdomwww.atlantic-chemicals.com CA00659 see Narender et al. Ref. Synthesis,(2007) 3469- 3472

S-29 Prepared from P-29 with MsCl and base followed by LiOHCombi-Blocks, LLC 7949 Silverton Ave. Suite 915 San Diego, CA 92126 USAwww.combi-blocks.com SS-5210

P-29

S-30 Prepared from P-29 with BOC anhydride and base followed by LiOH

S-31 Prepared from P-29 with AcCl and base followed by LiOH

S-32 Prepared from P-29 with MeOCOCl and base followed by LiOH

TABLE 16 Commercially Available Isomeric Thiazole acid Scaffolds andEster Precursors. Scaffold Scaffold Available Structure ID # CommentSource or Reference. Order # Precursor ID #

S-33 Aurora Fine Chemicals LLC 7929 Silverton Ave. Suite 609 San Diego,CA, 92126 USA www.aurorafinechemicals.com A02.445.497

S-34 Kingsh Chemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing,210036 People's Republic of China http://www.kingshchem.com 42198

S-35 Kingsh Chemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing,210036 People's Republic of China http://www.kingshchem.com 42196

S-36 ZereneX Molecular Limited 12 Manningham Road Greater Manchester,BL3 5QS United Kingdom www.zereneX-molecular.com ZXA008039

S-37 Kingsh Chemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing,210036 People's Republic of China http://www.kingshchem.com 43103

S-38 Prepared from P-38 by ester hydrolysis with LiOH Matrix ScientificP O Box 25067 Columbia, SC, 29224-5067 USAhppt://www.matrixscientific.com 39519

P-38

S-39 Prepared from P-39 with BOC anhydride and base then LiOH KingshChemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing, 210036People's Republic of China http://www.kingshchem.com 42007

P-39

S-40 Prepared from P-39 with MsCl and base then LiOH

A variety of additional ortho Aryl-5-membered heteroaryl carboxylic acidring systems that share the imidazole scaffold geometry have beenreported or are commercially available, Table 16. For example, Oxazolescaffolds 47 can be derived from 2-amino-ketoester 2 by simple acylationand acid catalized cyclization, Route-1, Scheme 8, (PCT Int. Appl. 20080704905, 21 Feb. 2008). The disubstituted triazole scaffold 49 can beprepared via the [3+2]cycloaddition of an aroyl actetic ester 10 withp-methoxybenzyl azide followed by deprotection, Route-2, (PCT Int. Appl.2005 073192, 11 Aug. 2005). Reaction of hydrazone 50 with methylacrylate followed by oxidation has be used to prepare the trisubstitutedpyrazole 51, Route-3, (PCT Int. Appl. 2005 080380, 1 Sep. 2005). Another[3+2]cycloaddition reaction involving an aryl azide 52 and methylpropynate provides a simple entrance to N-aryl triazole 53, Route-4. Allthe scaffold acids 54-57 share a similar geometry between the carboxylicacid function and the substitute aryl ring that is defined by the5-membered scaffold.

Commercially available oxazole acid scaffolds S-43 to S-49 and esterprecursors P-49 and P-49 are listed in Table 17. Complimentary2-mercaptooxazole derived scaffolds S-49, S-50 to S-53 and esterprecursor P-49 are listed in Table 18. The miscellaneous azoles acidscaffolds S-54 to S-64 summarized in Scheme 10 and bicyclic azole acidscaffolds S-65 to S-67 listed in Table 19 or their ester precursors areeither commercially available or readily prepared as indicated.

TABLE 17 Commercially Available Oxazole Acid Scaffolds and EsterPrecursors. Scaffold Scaffold Available Structure ID # Comment Source orReference. Order # Precursor ID #

S-41 See S-41 Experimental Section Feifei Zhao F., et. al., J. Org.Chem. (2011) 76 (24), pp 10338-10344

S-42 Otava Building Blocks Otava 55 Ellerslie Avenue, Suite 524 Toronto,ON, M2N 1X9 Canada http://www.otavachemicals.com 1788860

S-43 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www.bestpharmatech.com PB21319

S-44 Combi-Blocks, LLC 7949 Silverton Avenue, Suite 915 San Diego, CA,92126 USA www.combi-blocks.com HI-1354

S-45 OChem Incorporation 9044 Buckingham Park Drive Des Plaines, IL,60016 USA www.ocheminc.com 220N307

S-46 HDH Pharma, Inc. 7030 Kit Creek Road, Ste 250 Research TrianglePark, NC, 27709 USA www.hdhpharma.com  14091

S-47 Prepared from P-47 with LiOH APAC Pharmaceutical, LLC 6851 Oak HallLane Suite 101 Columbia, MD, 21045 USA Email: sales@apacpharma.comwww.apacpharma.com  650892

P-47

S-48 Prepared from P-48 with LiOH Otava Building Blocks Otava 55Ellerslie Avenue, Suite 524 Toronto, ON, M2N 1X9 Canadahttp://www.otavachemicals.com 1788860

P-48

TABLE 18 2-mercapto-Oxazole Derived Scaffolds. Scaffold AvailableStructure Scaffold ID # Comment Source or Reference. Order # PrecursorID #

S-49 Prepared from P-49 by ester hydrolysis with LiOH Nanosyn Inc. 3100Central Expressway Santa Clara, CA www.nanosyn.com NSN21446

P-49

S-50 Prepared from Precursor P-49 by alkylation with Mel follow by LiOHSee S-50 experimental section

S-51 Prepared from S-50 by treatment with 1-eq. of MCPBA See S-51experimental section

S-52 Prepared from S-50 by treatment with 2-eq. of MCPBA See S-52experimental section

A variety of commercially available azole scaffolds are listed in Table19 and Table-20.

TABLE 19 Sources for representative Miscellaneous Azole Scaffolds.Scaffold Scaffold Structure ID # Comment Source or Reference.

S-53 ChemBridge Corporation 16981 Via Tazon, Suite G San Diego, CA,92127 USA www.chembridge.com

S-54 ACB Blocks Ltd P O Box 10 Moscow, 121609 Russia Phone:+7(495)761-4365

S-55 Aurora Fine Chemicals LLC 7929 Silverton Ave. Suite 609 San Diego,CA, 92126 USA www.aurorafinechemicals.com

S-56 Prepared from P-56 by ester hydrolysis with LiOH Biolecule 160 NewBoston Street Woburn, MA, 01801 USA www.biolecule.com

S-57 Prepared from P-57 by ester hydrolysis with LiOH Biolecule 160 NewBoston Street Woburn, MA, 01801 USA www.biolecule.com

S-58 Best PharmaTech, Inc. P O Box 59394 Schaumburg, IL, 60159 USAhttp://www. bestpharmatech.com

S-59 Matrix Scientific P O Box 25067 Columbia, SC 29224 USAhttp://www.matrixscientific.com 38658

S-60 Prepared by multistep sequence See S-60 experimental section

S-61 Prepared from P-61 by ester hydrolysis with LiOH Lanzhou ChonChemical Co. Ltd. D6, Guchengping Industrial Park Donggang Town LanzhouCity People's Republic of China Web: http://www.chonchem.com

S-62 May be repared from Ethyl 3- Acyl-pyruvate and an appropriate arylhydrazine followed purification and LiOH hydrolysis.

S-63 PCT Int. Appl., 200305187226

S-64 Prepared form S-64 by reaction with MsCl and base PCT Int. Appl.,200305187226 Scaffold Scaffold Order ID Structure ID # # AvailablePrecursor #

S-53 4042713

S-54 2PA-0059

S-55 A00.603.652

S-56 Bi536

P-56

S-57 Bi548

P-57

S-58 BP42828

S-59 38658

S-61 P20978

P-61

S-62

TABLE 20 Commerically Available Bicyclic Scaffolds. Scaffold AvailableStructure Scaffold ID # Comment Source or Reference. Order # PrecursorID #

S-65 Kingsh Chemicals Ltd. 27-201 Jinwan Garden, Zhongbao St. Nanjing,210036 People's Republic of China http://www.kingshchem.com  44507

S-66 Otava Building Blocks Otava 55 Ellerslie Avenue, Suite 524 Toronto,ON, M2N 1X9 Canada http://www.otavachemicals.com 1515836

S-67 Prepared from P-65 by ester hydroylsis with LiOH Kingsh ChemicalsLtd. 27-201 Jinwan Garden, Zhongbao St. Nanjing, 210036 People'sRepublic of China http://www.kingshchem.com  44245

P-65

Efficient and general routes to oxazoles have been reported and may beutilized to prepare a wide variety of oxazole acid scaffolds, Scheme 9.As indicated, 2-amino-Aroylproionic esters 1 can be accessed by severalprocedures. Reaction of of 1 with trifluoroacetic anhydride andcyclodehydration provides an efficient approach to2-substituted-oxazoles, Route 1, and thiazoles, Route-3 (Sanz-Cervera,2009). Using these procedures, the oxazole scaffolds S-68 to S-80 may bereadily prepared Scheme 11.

Using the construction method outlined in Scheme 1, one skilled in theart could reasonably construct a two dimensional combinatorial librarycomprised of HGM-1 to HGM-73 described in Tables 1-9 and scaffold acidsS-1 to S-80 described in Tables 11 to 20 and Scheme 10. A constructedlibrary of the 5,840 compounds derived from these building blocks wouldbe expected to display interesting and new kinase inhibition profilesand attractive physical properties as lead compounds, useful tools fordiscovering new activities, and potential therapeutic agents.

In order to demonstrate the feasibility and usefulness of preparing sucha library, the 2×10 library of new inhibitors described in was preparedby coupling of amines HGM-1 and HGM-35 with the indicated scaffolds,Table 21 (p 129 or 130). These compounds were profiled against a panelof 230 kinases (Nanosyn Inc.). The kinases inhibited by ≧50% at 5 uMconcentration by these 20 new inhibitors is compared to the previouslyobserved activity profiles for Inhibitors I-14 and I-15, Table 14. Thesedata validate the utility of the scaffold geometry described herein forthe discovery of novel kinase inhibition profiles.

Methods for Preparation of Kinase Targeted PET Imaging Probes:

Our hypothesis that this unique scaffold geometry is broadly applicableto the Type-II inhibition of kinases suggests that coupling of anestablished Hinge-Gatekeeper Motif (HGM) with this scaffold could conveya similar activity profile. An example of this is SFE-0006, where theHGM of Sorafenib is incorporated. Therefore it may be assumed thatmodification of the HGM from known or experimental inhibitors couldconvey a similar activity profile. Therefore, chemistries have beendesigned to allow adaptation of this scaffold geometry for theconstruction of PET imaging agents, Scheme 11.

Preparation of Scaffolds:

Although a wide variety of ortho-aryl-5-membered-heteroaryl carboxylicacid scaffolds can be made, the preparation of specific inhibitorsdescribed below use the scaffolds illustrated in Schemes 1 to 4. Thesescaffolds vary primarily in their 2-substituent which vary in theirsize, polarity, and ability to establish hydrogen-bonds. For example the2-trifluoromethyl-imidazole scaffold S-1 and 2-methylthio-imidazole S-15are more lipophilic while the 2-methylsulfinyl, 2-methylsulfonyl-imidazole scaffolds S-16 and S-17 are significantly more polar,Scheme 12.

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylic Acid(S-1)

4-Fluoro-phenyl glycine 1 (10 g, 66 mmol) was added to a 250 mL roundbottomed flask that contained trifluoroacetic anhydride (30 mL) andstirred for 2 h at room temperature. Upon formation of oxazolinone 2,the excess TFAA and TFA were removed in vacuo as an azeotrope withtoluene. The remaining yellow solid was dissolved in 750 mL dry tolueneand benzylcyanoformate (11 mL, 1.1 equiv, 72 mmol) was added via syringefollowed by the dropwise addition of tributylphosphine (17 mL, 1 equiv,66 mmol). CO₂ emission from the reaction was observed with an oilbubbler. The reaction was allowed to stir at room temperature for 12 hand then was concentrated to dryness under reduced pressure, dissolvedin EtOAc, and washed with sodium bicarbonate, 1 M HCl, and brine. Thecrude mixture was purified by silica gel chromatography and eluted witha gradient of 0-60% EtOAc/Hex to give 7 g of the desired imidazolebenzyl ester 3 (7.0 g, 19.2 mmol) as an off-white solid.

Benzyl 5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylate3 (7.0 g, 19.2 mmol) was dissolved in ethanol (200 mL) and Pd-c (5%, 200mg) was added and the mixture was stirred under H₂(g) at 50 PSI on aParr shaker overnight. The mixture was filtered through celite and thesolvent was evaporated to give the desired product S-1 (5.25 g, 10mmol).

5-(4-Fluoro-phenyl)-2-methylthio-1H-imidazole-4-carboxylic acid (S-15)3-(4-Fluoro-phenyl)-2-hydroxyimino-3-oxo-propionic acid methyl ester (5)

To a stirred solution of methyl 4-fluorobenzoyl acetate (15.7 g, 80.0mmol) in HOAc (50 mL) was added a solution of NaNO₂ (8.28 g, 120 mmol)in water (80 mL) at 10° C. The resulting mixture was stirred at 10° C.for 1 h, then 16 h at rt. The precipitate formed in the reaction mixturewas collected by filtration, washed with water and dried over vacuum toafford product 5 (18.0 g, 100%) as a white solid. ESI MS m/z 240.2(M+H)⁺.

2-Amino-3-(4-fluoro-phenyl)-3-oxo-propionic acid methyl esterhydrochloride (6)

A solution of oxime 5 (18.0 g, 80.0 mmol) in MeOH (220 mL) was mixedwith 1.25 M HCl solution in MeOH (130 mL) at rt. The solution wasflushed with N₂ and then charged with 5% Pd—C (3.1 g). A hydrogenballoon was attached to the flask and the resulting mixture was stirredat rt for 2.5 h. More 5% Pd—C (˜1.8 g every 2 h) was added as thereaction was carefully monitored by LCMS. Eventually, the reaction wascompleted in 8 h with totally 9.4 g of 5% Pd—C added. After Pd catalystwas removed by filtering through a Celite pad, the clear solution wasconcentrated under reduced pressure to afford hydrochloride salt of pureproduct 6 (20.0 g, 100%) as a white solid. ESI MS m/z 212.2 (M+H)⁺.

5-(4-Fluoro-phenyl)-2-thioxo-2,3-dihydro-1H-imidazole-4-carboxylic acidmethyl ester (7)

To a stirred solution of aminoketone ester 6 hydrochloride (20.0 g, 80.0mmol) in DMF (100 mL) was added KSCN (15.6 g, 160 mmol) at rt. Theresulting mixture was stirred at 95° C. for 3 h and then allowed to coolto rt. After the reaction mixture was poured into brine (200 mL), awhite precipitate formed. Water (200 mL) was added to ensure the entireproduct 7 precipitated out from the resulting mixture. The precipitatewas collected by filtration, washed with water and dried on vacuum toafford product 7 (18.0 g, 89%) as a white solid. ESI MS m/z 253.2(M+H)⁺.

5-(4-Fluoro-phenyl)-2-methylsulfanyl-1H-imidazole-4-carboxylic acidmethyl ester (8)

To a stirred solution of compound 7 (18 g, 71.0 mmol) in MeOH (200 mL)was added K₂CO₃ (12.0 g, 85.0 mmol), followed by the dropwise additionof a solution of MeI (8.9 mL, 143 mmol) in MeOH (200 mL) at rt. Theresulting mixture was stirred at rt for 2 h and then concentrated underreduced pressure. The residue was treated with water (200 mL) andextracted with ethyl acetate (150 mL×3). The combined extracts werewashed with brine (50 mL×2), dried over anhydrous MgSO₄, filtered, andevaporated to afford product 8 (19.0 g, 100%) as a white solid. ESI MSm/z 267.2 (M+H)⁺.

5-(4-Fluoro-phenyl)-2-methylthio-1H-imidazole-4-carboxylic acid (S-15)

To a solution of 2-methylthioimidazole ester 8 (1 g, 3.6 mmol) indioxane (10 mL) was added 2M lithium hydroxide (18 mL, 36 mmol). Oxygenwas evacuated under vacuum and flask was flushed with nitrogen threetimes. Reaction mixture was stirred over night at 60° C. under nitrogen,brought to room temperature and gradually acidified with 1N HCl. Theresulting mixture was diluted with water and extracted with ethylacetate. Organic layer was washed with brine, dried with anhydroussodium sulfate, filtered and concentrated in vacuo to give 0.9 g(quantitave yield) of the desired product S-15.

5-(4-Fluoro-phenyl)-2-methylsulfinyl-1H-imidazole-4-carboxylic acid(S-16)

A solution of m-chloroperoxybenzoic acid (MCPBA) (579 mg, 3.35 mmol) in5 mL of dichloromethane was added drop wise to a stirred solution of 8(940 mg, 3.35 mmol) in 5 mL of dichloromethane at room temperature. Thereaction mixture was stirred for 4 h, and then transferred to aseperatory funnel and washed with saturated aq. NaHCO₃ solution, waterand brine. The organic layer was dried with anhydrous MgSO₄, filteredand concentrated in vacuo to give 1 g of the desired sulfoxide 9. Thiscrud sulfoxide intermediate was dissolved in 10 mL of dioxane and to itwas added 2M lithium hydroxide (18 mL, 36 mmol). Oxygen was evacuatedunder vacuum and flask was flushed with nitrogen three times. Reactionmixture was then stirred over night at 60° C. under nitrogen. Thereaction mixture was then gradually acidified with 1N HCl, diluted withwater and extracted with ethyl acetate. Organic layer was washed withbrine, dried with anhydrous sodium sulfate, filtered and concentrated invacuo to give 0.75 g (79% over two steps) of the product5-(4-Fluoro-phenyl)-2-methylsulfinyl-1H-imidazole-4-carboxylic acidS-16.

5-(4-Fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylic acid(S-17) 5-(4-Fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylicacid methyl ester (10)

To a stirred solution of compound 8 (19.3 g, 72.5 mmol) in MeOH (500 mL)was added a solution of OXONE (133 g) in water (500 mL) dropwise at rtin 1 h. The resulting mixture was stirred at rt for 4 h and thenconcentrated under reduced pressure. The slurry aqueous mixture wasextracted with DCM (200 mL×4). The combined extracts were washed withbrine (100 mL×2), dried over anhydrous MgSO₄, filtered, and evaporatedto afford sulfone 10 (19.8 g, 92%) as a white solid. ESI MS m/z 299.2(M+H)+.

5-(4-Fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylic acid

The sulfone 10 (19.8 g, 66.4 mmol) was treated with a mixture of LiOH(4.8 g, 200 mmol) in THF (240 mL) and water (80 mL) at 85° C. for 3 h.The resulting mixture was cooled to rt and then concentrated underreduced pressure. The aqueous solution was diluted with water (150 mL)and extracted with ethyl acetate (50 mL×2, discarded). The aqueoussolution was acidified with 1N HCl aq. to pH 1 and then extracted withethyl acetate (150 mL×3). The combined extracts were washed with brine(50 mL×2), dried over anhydrous MgSO₄, filtered, and evaporated toafford crude product 7 (18.0 g) as a white solid. ESI MS m/z 285.2(M+H)⁺. The crude product was suspended in a mixture of MTBE (50 mL) anddichloroethane (200 mL) and stirred at rt for 72 h. The remaining solidwas collected by filtration, dried over vacuum to afford pure5-(4-Fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylic acid(S-17) as a white solid (14 g, 74%) with 96% purity by UV254 nm and 100%purity by ELS.

5-(4-fluorophenyl)-2-(methylthio)thiazole-4-carboxylic acid (S-25) Ethyl5-(4-fluorophenyl)-2-(methylthio)thiazole-4-carboxylate (P-25)

Sodium thiomethoxide (2.13 g, 30.4 mmol) was added in three portionsover a period of 4 h to a solution of 2-Bromo-thiazole ester 11 (4 g,12.6 mmol) in dimethoxyethane (50 mL). Reaction mixture was then stirredovernight at room temperature. Solvent was removed under reducedpressure and the residue was suspended in water, filtered, washed withsmall amount of ethyl acetate and dried under high vacuum to afford 2.6g (72%) of 2-methylthio-thiazole ester P-25.

5-(4-fluorophenyl)-2-(methylthio)thiazole-4-carboxylic acid (S-25)

To a solution of ethyl5-(4-fluorophenyl)-2-(methylthio)thiazole-4-carboxylate 2 (2.6 g, 9.1mmol) in dioxane (20 mL) was added lithium hydroxide (0.32 g, 13.7 mmol)and the resulting mixture was stirred at 50° C. for two hours. Reactionmixture was then gradually acidified with 1N HCl. The precipitates werefiltered, washed with water and dried under high vacuum to give 2.4 g ofproduct S-25 (95%). ¹HNMR (CD₃OD, 250 MHz) δ 2.75 (s, 3H), 7.12 (t, 2H,J=8.75), 7.50 (dd, 2H, J=5.25, 8.75).

5-(4-fluorophenyl)-2-(methylsulfinyl)thiazole-4-carboxylic acid (S-26)

To a solution of S-25 (827 mg, 3 mmol) in dichloromethane (5 mL) wasadded m-CPBA (327 mg, 1.9 mmol) and the reaction mixture was stirred atroom temperature for 10 h. Reaction mixture was then evaporated todryness. To this residue diethylether was added to dissolve remainingm-CPBA and m-CBA. The suspended product was collected by filtration,washed with diethylether and dried under high vacuum to give 0.3 g (34%)of S-26. ¹HNMR (CD₃OD, 250 MHz) δ 3.12 (s, 3H), 7.17 (t, 2H, J=8.75),7.59 (dd, 2H, J=5.25, 8.75).

5-(4-fluorophenyl)-2-(methylsulfonyl)thiazole-4-carboxylic acid (S-27)

m-CPBA (516 mg, 3.3 mmol) was added to a stirred suspension of S-25 (300mg, 1.1 mmol) in dichloromethane (6 mL) and stirred overnight. LCMSanalysis showed some presence of sulfoxide. 86 mg (0.55 mmol) of m-CPBAwas added and the reaction mixture was further stirred for 2 hours.Reaction mixture was then evaporated to dryness and partitioned betweenethyl acetate and water. Organic layer was washed with brine, dried withanhydrous sodium sulfate, filtered and evaporated to dryness in rotaryevaporator to afford S-27 as crude product which is used directly in thenext step. ¹HNMR (CD₃OD, 250 MHz) δ 3.45 (s, 3H), 7.19 (t, 2H, J=9),7.62 (dd, 2H, J=5.25, 9).

5-(4-fluorophenyl)-2-(methylsulfonylamino)thiazole-4-carboxylic acid(S-29)

To a solution of P-29 (1 g, 3.96 mmol) in pyridine (10 mL) was addedmethanesulfonyl chloride (0.62 mL, 7.92 mmol) and the resulting mixturewas stirred at 60° C. over night. Reaction mixture was then brought toroom temperature and pyridine was evaporated under reduced pressure. Theresidue was dissolved in ethyl acetate and washed with saturated aq.sodium bicarbonate, water and brine. Organic layer was dried withanhydrous sodium sulfate, filtered and concentrated. The residue waspurified by flash chromatography (hexanes/ethyl acetate) to afford 1 g(77%) of product 12.

To a stirred suspension of intermediate 2 (1 g, 3 mmol) was added 2MLiOH in dioxane (7.5 mL, 3 mmol) and the solution was stirred for 2 h at40° C. The reaction mixture was then gradually acidified with 1N HCl.Diluted with water and extracted with ethyl acetate. Organic layer waswashed with brine and dried with anhydrous sodium sulfate. Filtrationand evaporation of organic layer afforded 0.9 g (94%) of the productS-29.

4-(4-Fluorophenyl)-2-(trifluoromethyl)oxazole-5-carboxylic acid. (S-41)(Z)-methyl 3-amino-3-(4-fluorophenyl)acrylate (13)

A MeOH (10 mL) solution of methyl 3-(4-fluorophenyl)-3-oxopropanoate(200 mg, 1.02 mmol) and NH₄OAc (393 mg, 5.10 mmol) was heated at refluxfor 18 hrs. Solvent was removed in vacuo and the product isolated bySiO₂ chromatography (70% hexanes/30% DCM to 100% DCM) product elutes in40% DCM (140 mg, colorless oil). 1HNMR (400 MHz, CDCl₃): d 7.46 (m, 2H),7.03 (m, 2H), 4.85 (s, 1H), 3.64 (s, 3H). 19FNMR (400 MHz, CDCl₃): d−115.3

Methyl 4-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-5-carboxylate (14)

To a solution of crude (Z)-methyl 3-amino-3-(4-flurophenyl)acrylate (100mg, 0.51 mmol) in dry DCE (5.1 mL) was added PhI(OCOCF₃)₂ (264 mg, 0.61mmol) in one portion at 45° C. The reaction mixture was heated at 45° C.for 3 hours as the reaction was monitored by TLC. The solvent wasevaporated and the crude material was separated by SiO₂ chromatography(Hexanes/DCM gradient from 80% hexanes/20% DCM to 100% DCM, desiredproduct elutes in 70% hexanes/30% DCM) to yield a white solid (49 mg,33% yield). 1HNMR (400 MHz, CDCl₃): d 8.15 (bs, 2H), 7.15 (m, 2H), 3.97(s, 3H). 19FNMR (400 MHz, CDCl₃): d −66.53, −109.59. MS(ESI): m/z 290.0[M+H]+

4-(4-Fluorophenyl)-2-(trifluoromethyl)oxazole-5-carboxylic acid (S-41)

To a solution of Methyl4-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-5-carboxylate (49 mg, 0.17mmol) in THF (1.5 mL) was added LiOH (21 mg, 0.51 mmol) in water (0.4mL). After 2 hours, the THF was evaporated, and the aqueous layer wasacidified with 1 N HCl and then extracted in EtOAc (3×15 mL). Theorganic fractions were dried over Na₂SO₄ and concentrated. 1HNMR (400MHz, MeOD/CDCl₃): d 7.99 (m, 2H), 7.03 (t, 2H, J=8.4 Hz). MS(ESI): m/z274.0 [M−H]− (S-41)

5-(4-fluorophenyl)-2-(methylthio)oxazole-4-carboxylic acid (S-50)

To a solution of P-49 (4 g, 15 mmol) in acetonitrile (30 mL) was addedmethyl iodide (1.4 mL, 22.5 mmol) and potassium carbonate (2 g, 15 mmol)and the resulting mixture was stirred at room temperature for 3 h.Reaction mixture was then diluted with water and extracted with ethylacetate. Organic layer was washed with brine, dried with anhydroussodium sulfate, filtered and concentrated in vacuo to give 4 g (95%) ofthe product 15.

To a solution of 1 (200 mg, 0.71 mmol) in dioxane (1 mL) was added 2Mlithium hydroxide (5 mL, 10 mmol). Oxygen was evacuated under vacuum andflask was flushed with nitrogen three times. Reaction mixture was thenstirred over night at 50° C. under nitrogen. The reaction mixture wasthen gradually acidified with 1N HCl, diluted with water and extractedwith ethyl acetate. Organic layer was washed with brine, dried withanhydrous sodium sulfate, filtered and concentrated in vacuo to give0.14 g (72%) of 5-(4-fluorophenyl)-2-(methylthio)oxazole-4-carboxylicacid S-50.

5-(4-fluorophenyl)-2-(methylsulfinyl)ozazole-4-carboxylic acid (S-51)

A solution of MCPBA (1.36 g, 7.9 mmol) in 10 mL of dichloromethane wasadded drop wise to a solution of S-50 (2 g, 7.9 mmol) in 10 mL ofdichloromethane. Reaction mixture was stirred for 4 h. Solvent was thenevaporated under reduced pressure and the product was purified by flashchromatography (MeOH/DCM) to give 1.57 g (73%) of sulfoxide S-51.

5-(4-fluorophenyl)-2-(methylsulfonyl)ozazole-4-carboxylic acid (S-52)

A solution of MCPBA (2.66 g, 15.4 mmol) solution in 10 mL ofdichloromethane was added drop wise to a solution of S-50 (1.95 g, 7.7mmol) in 10 mL of dichloromethane. Reaction mixture was stirredovernight at room temperature. Solvent was then evaporated under reducedpressure and the product was purified by flash chromatography (MeOH/DCM)to give 1.08 g (49%) of sulfone S-52.

5-(4-fluorophenyl)-2-(methylsulfonyl)-1,2-pyrazole-4-carboxylic acid(S-60)

Semicarbazide hydrochloride 17 (16.8 g, 151 mmol) and potassium acetate(16.8 g, 17 mmol) were suspended in ethanol (170 mL) and refluxed for 1h. The suspension was then filtered hot and 4-fluoroacetophenone 16 (20g, 144.7 mmol) was added to the mother liquor under reflux. Reactionmixture was stirred under reflux for another hour then cooled to roomtemperature and stirred overnight. Formed precipitates were filteredoff, washed with small amount of ethanol and vacuum dried to afford 24 g(87%) of product 18.

Phosphoryl chloride (2.1 mL, 23 mmol) was added drop wise to DMF (4.4mL, 57 mmol) at 0° C. The resulting mixture was kept on stirring for 30min at 0° C. and then 30 min at room temperature. Reaction mixture wascooled in ice/water bath and 18 (1.96 g, 10 mmol) was added slowly (notto exceed the temperature above 50° C.) then heated to 80° C. andstirred for 2 h at this temperature. Reaction mixture was then pouredinto ice (15 g), basified with 5N NaOH till pH 10 and stirred for 2 hthen acidified with concentrated HCl and continued to stir foradditional 4 h. Formed precipitates were filtered off, washed with smallamount of water and dried under high vacuum to afford 1.5 g (79%) ofaldehyde 19.

Methanesulfonyl chloride (5.4 mL, 70 mmol) was added to a solution of 17(4 g, 21 mmol) in pyridine (50 mL) and stirred at 60° C. for 10 h.Additional 3 equivalents of MsCl (5.4 mL, 70 mmol) were added and thereaction was continued to stir at 60° C. for further 12 h. Reactionmixture was then brought to room temperature and pyridine was removedunder reduced pressure. Residue was dissolved in ethyl acetate andwashed carefully with saturated aq. sodium bicarbonate, water and brine.Organic layer was dried with anhydrous sodium sulfate, filtered andconcentrated to give 3 g (54%) of product 20.

To a solution of 18 (1.75 g, 6.5 mmol) in DMF (50 mL) was added oxone(4.8 g, 7.8 mmol) and the resulting mixture was stirred overnight atroom temperature. DMF was removed under reduced pressure and the residuewas partitioned between ethyl acetate and water. Organic layerseparated, washed with brine, dried with anhydrous sodium sulfate andconcentrated in vacuo to afford 1.5 g (81%) of pyrazole acid S-60.

Ethyl 5-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-4-carboxylate (S-68)Ethyl 3-(4-F-phenyl)-3-oxo-2-(2,2,2-trifluoroacetamido)propanoate (21)

To a solution of ethyl 2-amino-3-(4-fluorophenyl)-3-oxopropanoate (50mg, 0.22 mmol) and pyridine (105 uL, 1.34 mmol) in toluene (2.5 mL) andDCM (0.5 mL) on ice was added trifluoroacetic anhydride (78 mL, 0.56mmol). The reaction was warmed to room temperature and allowed to stirovernight. The major product was isolated by SiO2 chromatography(isocratic DCM) to afford a colorless oil (37 mg, 51% yield). 1HNMR (400MHz, CDCl3): d 8.12 (m, 2H), 7.68 (bs, 1H), 7.15 (t, 2H, J=8.4 Hz), 6.03(d, 1H, J=6.8 Hz), 4.14 (m, 2H), 1.10 (t, 3H, J=6.8 Hz). 19FNMR (400MHz, CDCl3): d −76.10, −101.37. MS(ESI): m/z 321.9 [M+H]+

Ethyl 5-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-4-carboxylate (22)

Triethylamine (64 mL, 0.46 mmol)) was added to a solution oftriphenylphosphine (60 mg, 0.23 mmol) and iodine (58 mg, 0.23 mmol) indry DCM (0.5 mL) and stirred for 5 min. Then a solution of ethyl3-(4-fluorophenyl)-3-oxo-2-(2,2,2-trifluoroacetamido)propanoate (37 mg,12 mmol) in dry DCM (0.7 mL) was added and the reaction mixture wasstirred for 1 hr. The solvent was removed under reduced pressure, andthe residue was purified by SiO2 chromatography on silica gel (isocraticDCM) to yield a white solid (31 mg, 88% yield). 1HNMR (400 MHz, CDCl3):d 8.12 (m, 2H), 7.18 (t, 2H, J=8.8 Hz), 4.43 (q, 2H, J=7.2 Hz), 1.39 (t,3H, J=7.2 Hz). 19FNMR (400 MHz, CDCl3): d −66.10, −107.21. MS(ESI): m/zMS(ESI): m/z 303.9 [M+H]+

5-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-4-carboxylic acid (23)

Compound ethyl5-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-4-carboxylate (150 mg,0.49 mmol) was dissolved in THF (4.6 mL) and cooled on ice. To thissolution is added LiOH (13 mg, 0.31 mmol) in water (1.5 mL) and themixture was stirred for 1 hr then warmed to room temperature and stirredovernight. The THF was evaporated, and H2O (10 mL) was added and thesolution then acidified with 1 N HCl and extracted in EtOAc (3×15 mL).The organic fractions were dried over Na₂SO₄ and concentrated. The crudematerial was fractionated by SiO2 chromatography (DCM to 90% DCM/10%MeOH gradient, the desired product elutes in 7% MeOH) to afford a whitesolid (17 mg, 11% yield). 1HNMR (400 MHz, MeOD, CDCl₃): d 8.17 (m, 2H),7.14 (t, 2H, J=8 Hz).

Ethyl 5-(4-fluorophenyl)-2-(trifluoromethyl)thiazole-4-carboxylate(S-75)

To a solution of ethyl3-(4-fluorophenyl)-3-oxo-2-(2,2,2-trifluoroacetamido)propanoate (271 mg,0.84 mmol) in anhydrous THF (10 mL) was added Lawesson's reagent (682mg, 1.69 mmol) in one portion and the resulting solution was refluxedfor 22 hrs under nitrogen until no starting material is detected by TLC.The solvent was removed under reduced pressure and the crude materialwas purified by SiO₂ chromatography (80% hexanes/20% DCM to 100% DCM ina linear gradient, product elutes in 50% hexanes/50% DCM) (175 mg, 64%yield). 1HNMR (400 MHz, CDCl₃): d 7.50 (m, 2H), 7.15 (t, 2H, J=8.8 Hz),4.31 (q, 2H, J=6.8 Hz), 1.26 (t, 3H, 7.2 Hz).

2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-1H-imidazole-4-carboxylicacid, (S-81) Step-1: ethyl 5-(4-fluorophenyl)oxazole-4-carboxylate

Using the procedure described in Russian Journal of Organic Chemistry 42(7), 1031-1035 (2006), ethyl 5-(4-fluorophenyl)oxazole-4-carboxylate wasprepared as follows: Sodium hydride (42 mg, 1.04 mmol, 60% in oil) wassuspended in benzene (1 mL) and to this suspension was added ethylisocyanoacetate (102 mg, 0.90 mmol) in benzene (0.5 mL) and the mixturewas stirred until no more gas evolution was evident (1-2 h). To thismixture was added p-fluorobenzoyl chloride (165 mg, 1.04 mmole) inbenzene (0.5 mL). The resulting reaction mixture was stirred overnightat room temperature. The crude product was obtained by partitioningbetween ethyl acetate and water and evaporation to dryness. Purificationby flash chromatography afforded the title compound. LC/MS 236 M+H⁺

Step-2: ethyl 5-(4-fluorophenyl)-2-mercaptoimidazole-4-carboxylate

Using the procedure described in Chemical and Pharmaceutical Bulletin 32(7), 2536-45 (1984), ethyl5-(4-fluorophenyl)-2-mercaptoimidazole-4-carboxylate was prepared asfollows:

Ethyl 5-(4-fluorophenyl)oxazole-4-carboxylate (0.235 g, 1 mmol) wasdissolved in a mixture of concentrated HCl and ethanol (1:3) and themixture was heated at 50° C. for 4 h. The solvents were removed underreduced pressure and the crude product thoroughly dried. The crudehydrochloride was dissolved in water and treated with excess potassiumthiocyanate. The solution was heated at 80-90° C. for 4 h and thencooled in ice. The solution was filtered and the precipitaterecrystallized from ethanol to afford the title compound.

Step-3:2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-1H-imidazole-4-carboxylicacid

5-(4-Fluoro-phenyl)-2-mercapto-1H-imidazole-4-carboxylic acid ethylester (3 g, 11.28 mmol) was dissolved in EtOH (30 mL) at RT. K₂CO3 (6.23g, 45 mmol) and (2-chloro-ethyl)-dimethyl-amine HCl salt (1.54 g, 10.72mmol) were added. The mixture was stirred at 50° C. for 48 h. It wasthen concentrated and the residue was dissolved in THF/water (20 mLeach) at RT. NaOH (2 g, 50 mmol) was added. The mixture was stirred atRT for 12 h. It was then concentrated to remove most THF and the pH ofthe solution was adjusted to ˜5 using 3 N HCl. The mixture wasconcentrated and residue was purified by prep HPLC. Lyophilization ofpure fractions afforded 1.1 g pure2-(2-dimethylamino-ethylthio)-5-(4-fluorophenyl)-1H-imidazole-4-carboxylicacid, S-81. LC-MS [M+H] 310.2.

1-(4-fluorophenyl)-3-(methylthio)-1H-pyrazole-5-carboxylic acid, (S-81)

5-(4-Fluoro-phenyl)-2-mercapto-1H-imidazole-4-carboxylic acid ethylester (3 g, 11.28 mmol) was dissolved in EtOH (30 mL) at RT. K₂CO3 (6.23g, 45 mmol) and (2-chloro-ethyl)-dimethyl-amine HCl salt (1.54 g, 10.72mmol) were added. The mixture was stirred at 50° C. for 48 h. It wasthen concentrated and the residue was dissolved in THF/water (20 mLeach) at RT. NaOH (2 g, 50 mmol) was added. The mixture was stirred atRT for 12 h. It was then concentrated to remove most THF and the pH ofthe solution was adjusted to ˜5 using 3 N HCl. The mixture wasconcentrated and residue was purified by prep HPLC. Lyophilization ofpure fractions afforded 1.1 g pure2-(2-dimethylamino-ethylthio)-5-(4-fluorophenyl)-1H-imidazole-4-carboxylicacid. LC-MS [M+H] 310.2.

Preparation of 3-bromo-1-(4-fluorophenyl)-1H-pyrazole-5-carboxylic acid,(S-82)

5-Bromo-2-(4-fluoro-phenyl)-2H-pyrazole-3-carboxylic acid was preparedfollowing procedures from WO 2006055922, using 4-fluorophenylhydrazineinstead of 2-chlorophenylhydrazine.

Step-1: (E)-2-(2-(4-fluorophenyl)hydrazono)acetic acid

To a solution of 4-fluorophenyl hydrazine 1.32 g (0.011 mol) in water(30 mL) at room temperature was added HCl (1.32 g, 0.014 mol) followedby the drop-wise addition of glyoxylic acid (50%, 1.71 g, 0.012 mol)over 20 minutes. The thick suspension was stirred for 30 m. The productwas isolated by filtration, washed with water and then dissolved inethyl acetate (40 mL), dried over magnesium sulfate, filtered and theproduct isolated as a solid.

Step-2: (4-fluorophenyl)carbonohydrazonic dibromide

To the crude product of the previous step 1.88 g (0.010 mol) dissolvedin DMF (20 mL) at 0° C. was added N-bromosuccinimide (3.57 g, 0.021 mol)in portions over 30 m. The resulting mixture was stirred overnight atroom temperature. The mixture was partitioned between ether and water(20:15) and extracted twice more with ether (20 mL). The combinedorganic extracts were evaporated and purified by flash chromatography toafford the title compound.

Step-3: methyl3-bromo-1-(4-fluorophenyl)-4,5-dihydro-1H-pyrazole-5-carboxylate

To the crude product obtained in step 2, 1.14 g (3.85 mmol) in DMF (12mL) was added methyl acrylate (1.39 mL, 15.4 mmol) all at once.Diisopropylethylamine (0.74 mL, 4.23 mmol) was added dropwise over 15 m.The mixture was stirred at room temperature for 1 h and then dilutedwith water and extracted twice with ether. Concentration of the solventafter drying over magnesium sulfate afforded the title compound.

Step-4: methyl 3-bromo-1-(4-fluorophenyl)-1H-pyrazole-5-carboxylate

The product obtained in step 3 1.15 g (3.84 mmol) was dissolved inacetone (40 mL) and treated with potassium permanganate (2.42 g, 15.4mmol) in portions of 100 mg at 10 minute intervals taking care to keepthe reaction temperature below 40° C. The mixture was stirred overnightat room temperature. Filtration through Celite removed the insolublematter and the filter cake was washed with ether. Solvent was removedand the product purified by flash chromatography to afford the titleproduct. (0.58 g).

Step-5: 3-bromo-1-(4-fluorophenyl)-1H-pyrazole-5-carboxylic acid

The crude ester 0.58 g (1.84 mmol) obtained in the previous step washydrolyzed in methanol (5 mL) with aqueous sodium hydroxide (0.88 g in7.5 mL, 3.05 mmol). After stirring at room temperature for 2 h, themixture was diluted with water and extracted with ether. The aqueouslayer was acidified with HCl to pH 2 and the product extracted withtwice with ethyl acetate. The solution was dried and evaporated todryness to afford the pure acid, S-82 (0.54 g).

1-(4-fluorophenyl)-3-(methylthio)-1H-pyrazole-5-carboxylic acid, (S-83)

2-(4-Fluoro-phenyl)-5-methylthio-2H-pyrazole-3-carboxylic acid wasprepared following the procedure described in the literature (EuropeanJournal of Medicinal Chemistry, 43(8), p 1715, 2008 and ChemischeBerichte, 112(4), p 1206, 1979).

Step 1: Preparation of Ethyl N-(4-fluorophenyl) glycine

Using the procedure described in WO 9857937 A2, to a solution of 13.56 g(122 mmole) p-fluoroaniline in DMF (100 mL) under nitrogen was added23.5 g (141 mmol) of ethyl bromoacetate and 14.95 g (141 mmol) anhydroussodium carbonate. The mixture was heated to 70° C. for 16 h and thencooled to room temperature. Water was added (500 mL) and the mixturestirred vigorously until a precipitate formed. The solid was filtered,washed with 100 mL water and dried to afford ethyl N-(4-fluorophenyl)glycine.

Step-2: Preparation of N-(4-fluorophenyl) glycine

To a solution of 21.3 g (108 mmole) of ethyl N-(4-fluorophenyl) glycinein THF (100 mL) under nitrogen was added LiOH (5.44 g (130 mmol)monohydrate in 25 mL water. After 15 h the mixture was concentrated tohalf the volume and acidified to pH 3 with HCl. The precipitated solidwas collected and washed with water (100 mL) to afford the titlecompound.

Step-3: Preparation of N-(4-fluorophenyl)-N-nitrosoglycine

Sodium nitrite (3.97 g), 57.5 mmol) in 10 mL water was added to asuspension of N-(4-fluorophenyl) glycine 9.25 g (54.7 mmol) in water (50mL) under nitrogen. The reaction mixture was stirred until clear, ca. 6hours. Acidification to pH 3 with HCl precipitated the product which wasfiltered and washed with water (50 mL) and dried to afford the titleproduct.

Step-4: Preparation of3-(4-fluorophenyl)-3H-1,2,3-oxadiazol-1-ium-5-olate

N-(4-fluorophenyl)-N-nitrosoglycine (11.5 g, 54.7 mmol) was dissolved inacetic anhydride (100 mL) and heated to 70° C. for 14 h. The reactionmixture was cooled and then poured into ice water (300 mL) After forstirring for 30 m, the reaction mixture was filtered to provide 10.50 g(100%) of the title product.

Step 5—Preparation of3-(4-fluorophenyl)-4-(methylthio)-3H-1,2,3-oxadiazol-1-ium-5-olate

Using the procedure of Masuda and Okutani (Tetrahedron 30, 409-414(1974)) the crude 3-(4-fluorophenyl)-3H-1,2,3-oxadiazol-1-ium-5-olate(1.95 g, 10.82 mmol) was dissolved in anhydrous DMSO (27 mL) and cooledin ice. Acetyl chloride (2.0 g, 25.5 mmol) was added dropwise. Thereaction mixture was allowed to come to room temperature. After stirringovernight, the mixture was partitioned between ether and saturatedNaHCO₃ solution. The organic layer was washed several times with water,dried and evaporated to dryness to afford3-(4-fluorophenyl)-4-(methylthio)-3H-1,2,3-oxadiazol-1-ium-5-olate.

Step 6—Preparation of1-(4-fluorophenyl)-3-methylthio-pyrazole-5-carboxylate

1-(4-fluorophenyl)-3-methylthio-pyrazole-5-carboxylate was prepared bythe cycloaddition—rearrangement reaction described in ChemischeBerichte, 112(4), 1206 (1979). Thus crude3-(4-fluorophenyl)-4-(methylthio)-3H-1,2,3-oxadiazol-1-ium-5-olate,(1.73 g, 7.68 mmole) and methyl propiolate (6.45 g, 76.8 mmole) weredissolved in CH₂Cl₂ (10 mL) and the quartz reaction vessel purged withnitrogen. The reaction mixture was irradiated in a Rayonet RPR-100photochemical reactor for 14 h. The crude product was concentrated invacuo and then purified by flash chromatography to afford methyl1-(4-fluorophenyl)-3-methylthio-pyrazole-5-carboxylate, 374 mg afterflash chromatography.

Step 7—Preparation of1-(4-fluorophenyl)-3-(methylthio)-1H-pyrazole-5-carboxylic acid

The crude ester 0.374 g (1.40 mmol) obtained in the previous step washydrolyzed in methanol (5 mL) with aqueous sodium hydroxide (0.67 g in7.5 mL, 2.32 mmol). After stirring at room temperature for 2 h, themixture was diluted with water and extracted with ether. The aqueouslayer was acidified with HCl to pH 2 and the product extracted withtwice with ethyl acetate. The solution was dried and evaporated todryness to afford the pure acid S-83.

1-(4-fluorophenyl)-3-(methylsulfonyl)-1H-pyrazole-5-carboxylic acid,(S-84)

1-(4-fluorophenyl)-3-(methylthio)-1H-pyrazole-5-carboxylic acid (120 mg,0.48 mmol) was dissolved in acetonitrile (5 mL) at RT. Oxone™ (440 mg,1.4 mmol) was added. The mixture was stirred at RT for 30 min and thenheated at 50° C. for 3 h. The mixture was cooled to RT and mostvolatiles were removed under vacuum. The residue was partitioned betweenDCM/brine. The DCM layer was dried over Na₂SO₄ and filtered.Concentration of the filtrate afforded the crude1-(4-fluorophenyl)-3-(methylsulfonyl)-1H-pyrazole-5-carboxylic acid(˜140 mg), which was used without further purification. LC-MS [M+H]285.1.

Preparation of Inhibitors:

2-Chloro-5-nitro-benzoic acid methyl ester (15)

2-chloro-5-nitrobenzoic acid 14 (5.0 g, 24.8 mmol) was dissolved inmethanol (10 mL) and 4 N HCl in dioxane (12 mL) was added. The mixturewas stirred for 3 hr after which the solvents were evaporated and theresidue was neutralized and extracted with EtOAc, washed with NaHCO₃,brine, dried over MgSO₄ and filtered. The solvent was evaporated to givethe desired product 15 (3.9 g, 75%).

5-Amino-2-chloro-benzoic acid methyl ester (16)

2-Chloro-5-nitro-benzoic acid methyl ester 15 (4.3 g, 20.2 mmol),SnCl₂-2H₂O (45.5 g, 201.8 mmol) and ethanol (100 mL) were combined intoa sealed pressure vial and was sonicated for 2 hr. The solvent wasconcentrated and the residue was dissolved in EtOAc, washed with NaHCO₃,brine, dried over MgSO₄ and filtered. The solvent was evaporated to givethe desired product 16 (2.8 g, 73%).

Methyl2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoiate(17)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylic acid 16(2.0 g, 7.3 mmol) was dissolved in DCM (25 mL). To the mixture was addedHATU (3.0 g, 7.95 mmol) and DIEA (3.5 mL, 19.9 mmol) after 15 min ofstirring, 5-Amino-2-chloro-benzoic acid methyl ester (1.2 g, 6.6 mmol)was added. The reaction was refluxed for 48 hr. The solvent wasevaporated and the residue was purified by silica gel chromatography(EtOAc/Hx, 0-40%) to yield 0.93 g (33%) of the desired product 17.

2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoicacid (18)

Methyl2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoate-17(0.93 g) was dissolved in water (4.2 mL) and LiOH (177.2 mg, 4.2 mmol)was added and the mixture was stirred for 2 hr. Acetic acid (gl., 0.24mL) was added. The resulting precipitated was filtered and washed withwater. The solid was dissolved in EtOAc, dried over MgSO₄, filtered andthe solvent was evaporated to give 1.0 g of the crude product 18 whichwas used without further purification.

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid[4-chloro-3-(pyridin-2-ylcarbamoyl)-phenyl]-amide—(I-1)

2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoicacid 18 (0.2 g, 0.42 mmol) was dissolved in DCM (2 mL) and HATU (0.32 g,0.84 mmol) and DIEA (0.16 mL, 0.94 mmol) after 15 min of stirring,2-Aminopyridine (0.053 g, (1.2 g, 0.56 mmol) was added and the mixturewas stirred overnight. The mixture was washed with NaHCO₃, water, brine.The organic layer was dried over MgSO₄, filtered and evaporated. Theresidue was purified by HPLC to give only 1.7 mg of the desired product(I-1).

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid[4-chloro-3-(pyridin-4-ylcarbamoyl)-phenyl]-amide—(I-2)

2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoicacid 18 (0.2 g, 0.42 mmol) was dissolved in DCM (2 mL) and HATU (0.32 g,0.84 mmol) and DIEA (0.16 mL, 0.94 mmol) after 15 min of stirring,4-Aminopyridine (0.053 g, (1.2 g, 0.56 mmol) was added and the mixturewas stirred overnight. The mixture was washed with NaHCO₃, water, brine.The organic layer was dried over MgSO₄, filtered and evaporated. Theresidue was purified by HPLC to give 17.5 mg of the desired product(I-2).

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid[4-chloro-3-(pyridin-3-ylcarbamoyl)-phenyl]-amide—(I-3)

2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoicacid 18 (0.2 g, 0.42 mmol) was dissolved in DCM (2 mL) and HATU (0.32 g,0.84 mmol) and DIEA (0.16 mL, 0.94 mmol) after 15 min of stirring,3-Aminopyridine (0.053 g, (1.2 g, 0.56 mmol) was added and the mixturewas stirred overnight. The mixture was washed with NaHCO₃, water, brine.The organic layer was dried over MgSO₄, filtered and evaporated. Theresidue was purified by HPLC to give 95.7 mg of the desired product(I-3).

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid[3-(3-amino-pyrazin-2-yloxymethyl)-4-chloro-phenyl]-amide—(I-8)

2-Chloro-5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-benzoicacid methyl ester 17 (0.165 g, 0.38 mmol) was dissolved in DCM (0.75 mL)and cooled to −78° C. To the mixture was added DiBAL-H (1M, 1.2 mL, 1.12mmol) and the reaction was allowed to stir for 1 hr. The reaction wasquenched with methanol and treated with ammonium chloride (sat.). TheDCM layer was washed with water and dried over MgSO₄. The solvent wasfiltered and evaporated to provide the intermediate alcohol 19 (71 mg)which was used without further purification. (71 mg, 0.17 mmol) wasdissolved in DMSO and NaH (60%, 0.024 g, 0.51 mmol) was added underN₂(g). To the mixture was added 2-amino-3-chloropyrazine (0.023 g, 0.17mmol) and reaction was allowed to stir overnight. Water was added andthe sample was extracted with DCM and dried over MgSO₄. Afterevaporation, the residue was dissolved in DMSO and purified by HPLC togive 11.6 mg of the desired product (I-8).

(2-Methyl-5-nitro-phenyl)-carbamic acid tert-butyl ester (21)

2-methyl-5-nitroaniline 20 (5.0 g, 32.86 mmol) and BOC anhydride (8.6 g,39.44 mmol) were dissolved in dichloromethane (110 mL). Pyridine (5.4mL, 65.72 mmol) was added to the mixture and the reaction was stirredfor 16 hrs. The solvents were evaporated and the residue was purified bysilica gel chromatography (EtOAc/Hx, 0-50%) to yield 4.5 g (54%) of thedesired product 21.

(5-Amino-2-methyl-phenyl)-carbamic acid tert-butyl ester (22)

(2-Methyl-5-nitro-phenyl)-carbamic acid tert-butyl ester 21 (4.5 g, 17.8mmol) was dissolved in ethanol (100 mL) and Pd-c (5%, 100 mg) was addedand the mixture was stirred under H₂(g) at 50 PSI on a Parr shakerovernight. The mixture was filtered through celite and the solvent wasevaporated to give the desired product 22 (4.5 g, quantitative yield).

(5-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-carbamicacid tert-butyl ester (23)

(5-Amino-2-methyl-phenyl)-carbamic acid tert-butyl ester 22 (2.0 g, 7.3mmol) was dissolved in DCM (25 mL) and DIEA (3.8 mL, 21.8 mmol) wasadded. To the mixture was added HATU (3.3 g, 8.7 mmol), after 15 min ofstirring,5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylic acid S-1(2.0 g, 7.3 mmol) was added. The reaction was refluxed for 48 hr. Thesolvent was evaporated and the residue was purified by silica gelchromatography (EtOAc/Hx, 0-40%) to yield 2.5 g (72%) of the desiredproduct 23.

(5-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-amine(24)

(5-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-carbamicacid tert-butyl ester 23 (2.5 g, 5.2 mmol) was dissolved in a mixture ofDCM/TFA (50%) and stirred overnight. The solvents were evaporated andthe residue was neutralized and extracted with EtOAc, washed with brine,dried over MgSO₄, filtered and evaporated. The residue was purified bysilica gel chromatography (EtOAc/Hx, 0-75%) to yield 2.5 g (72%) of thedesired product 24 (1.6 g, 82%).

N-(5-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-nicotinamide—(I-4)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid and(3-amino-4-methyl-phenyl)-amide 6 (0.2 g, 0.53 mmol) was dissolved inDCM (2 mL) and DIEA was added to the solution (0.277 mL, 1.59 mmol).Nicotinyl chloride HCl (0.095 g, 0.53 mmol) was added and the mixturewas stirred overnight. The mixture was washed with NaHCO₃, water, brine.The organic layer was dried over MgSO₄, filtered and evaporated. Theresidue was purified by HPLC to give 160 mg of the desired product(I-4).

N-(5-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-isonicotinamide—(I-5)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid and(3-amino-4-methyl-phenyl)-amide 6 (0.2 g, 0.53 mmol) was dissolved inDCM (2 mL) and DIEA was added to the solution (0.277 mL, 1.59 mmol).Pyridine-4-carboxylic acidchloride HCl (0.095 g, 0.53 mmol) was addedand the mixture was stirred overnight. The mixture was washed withNaHCO₃, water, brine. The organic layer was dried over MgSO₄, filteredand evaporated. The residue was purified by HPLC to give 160 mg of thedesired product (I-5).

1H-Benzoimidazole-5-carboxylic acid(5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-amide—(I-9)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid and(3-amino-4-methyl-phenyl)-amide 6 (0.20 g, 0.53 mmol) was dissolved intoluene (1.0 mL). To the reaction was addedmethyl-1H-benzimidazole-5-carboyxlate (0.11 g, 0.63 mmol) and 2.0Mtetramethyl aluminum (0.50 mL, 1.03 mmol). The mixture was heated undermicrowave conditions for 20 min at 160° C. The solvent was evaporated;DMSO was added to the residue and filtered. The filtrate was purified byHPLC to give 2.2 mg of the desired product (I-9).

1H-Benzotriazole-5-carboxylic acid(5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-amide—(I-10)

1-H-Benzotriazole-5-carboxylic acid (0.19 g, 0.56 mmol) was dissolved ina mixture of DCM (2 mL) and DMF (1 mL). To the mixture was added HATU(0.21 g, 0.56 mmol) and DIEA (0.122 mL, 0.74 mmol) after 15 min ofstirring,5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid(3-amino-4-methyl-phenyl)-amide 6 (0.07 g g, 0.19 mmol) was added. Thereaction was stirred at rt for 48 hr. The solvent was concentrated todryness under reduced pressure, dissolved in EtOAc, and washed withsodium bicarbonate, 1 M HCl, and brine. The solvent was evaporated andthe residue was purified by HPLC to yield 28.4 mg of the desired product(I-10).

1H-7-Azaindole-3-carboxylic acid(5-{[5-(4-fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-2-methyl-phenyl)-amide—I-11

7-azaindole-3-carboxylic acid (0.19 g, 0.56 mmol) was dissolved in amixture of DCM (2 mL) and DMF (1 mL). To the mixture was added HATU(0.21 g, 0.56 mmol) and DIEA (0.122 mL, 0.74 mmol) after 15 min ofstirring,5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carboxylic acid(3-amino-4-methyl-phenyl)-amide 6 (0.07 g g, 0.19 mmol) was added. Thereaction was stirred at rt for 48 hr. The solvent was concentrated todryness under reduced pressure, dissolved in EtOAc, and washed withsodium bicarbonate, 1 M HCl, and brine. The solvent was evaporated andthe residue was purified by HPLC to yield 23.5 mg of the desired product(I-11).

The quinazolinone Hing-Gatekeeper anchor intermediate was prepared asoutlined in WO 2007 GB 1389; WO2008120004, Scheme-4

6-(5-Amino-2-methylphenylamino)-3-methylquinazolin-4(3H)-one—30:Tert-butyl-4-methyl-3-nitrophenylcarbamate

A solution of 4-methyl-3-nitroaniline 25 (15.0 g, 99 mmol) was dissolvedin 50 mL THF at 65° C. 25.8 g Boc anhydride was dissolved in 30 mL THFand added dropwise over the course of 20 minutes. The reaction wasrefluxed overnight, cooled to room temperature, and then concentrated invacuo to yield a brown oil. The oil was dissolved in 150 mL 20%EtOAc/Hex and 50 g of silica gel was added. The solution was stirred for15 min on a rotovap and the silica was removed by filtration and washedwith 300 mL 20% EtOAc/Hex. The organics were dried over magnesiumsulfate and then concentrated by vacuum filtration to yield 20.7 gproduct in 83% yield. ¹H NMR (300 MHz, D₆ DMSO): δ 9.75 (br s, 1H), 8.22(d, 1H, J=2.1 Hz), 7.58 (dd, 1H, J=2.1 Hz, J=8.3 Hz), 7.37 (d, 1H, J=8.3Hz), 2.43 (s, 3H), 1.48 (s, 9H). ¹³C NMR (75 MHz, D₆ DMSO): δ 153.50,148.48, 139.45, 133.79, 126.65, 123.52, 113.81, 80.60, 28.84, 19.92.ESMS: m/z 270.0 [M+H20].

Tert-butyl 3-amino-4-methylphenylcarbamate—26

Tert-butyl 4-methyl-3-nitrophenylcarbamate (20.7 g, 82.0 mmol) wasdissolved in 200 mL EtOH and 1.0 g of 5% Pd/C was added. The mixture washydrogenated on a parr system at 50 PSI hydrogen for 15 hours.Completion of reaction was shown by TLC in 30% EtOAc/Hex, then filtered,concentrated, and dried in a vacuum oven to yield the product 26 (16.78g, 75.0 mmol, 92% yield). ¹H NMR (300 MHz, D₆ DMSO): δ 8.93 (br s, 1H),6.85 (s, 1H), 6.75 (d, 1H, J=8.0 Hz), 6.5 (d, 1H, J=8.0 Hz), 4.75 (s,2H), 1.97 (s, 3H), 1.46 (s, 9H). ¹³C NMR (75 MHz, D₆ DMSO): δ 153.58,147.37, 138.777, 130.51, 115.87, 107.53, 105.07, 79.20, 29.05, 17.71.ESMS: m/z 223.2 [MH]⁺

6-Chloro-3-methylquinazolin-4(3H)-one 28

2-amino-5-chlorobenzoic acid 27 (12.5 g, 73 mmol) was placed in a 250 mLbeaker containing n-methylformamide (125 mL). A condenser was utilizedand the reaction was heated overnight at 180° C. The reaction wascomplete the next morning and allowed to cool to room temperature for 1hr and then poured into 500 mL of water and extracted three times with250 mL of ethyl acetate. The organic phase was then washed with anadditional 1 liter of water and concentrated under vacuum to yield6-chloro-3-methylquinazolin-4(3H)-one (9.2 g, 47.3 mmol, 65% yield)¹HNMR (300 MHz, D₆ DMSO): δ 8.39 (s, 1H), 8.04 (d, 1H, J=3.6 Hz), 7.82(dd, 1H, J=2.4 Hz, J=8.7 Hz), 7.67 (1H, d, J=8.7 Hz), 3.49 (s, 3H). ¹³CNMR (75 MHz, D₆ DMSO): δ 160.49, 149.75, 147.65, 135.06, 131.99, 130.26,125.61, 123.45, 34.53. ESMS: m/z 195.2 [MH]⁺

Tert-butyl4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenylcarbamate29

To refluxing toluene (50 mL) in a 150 mL round bottom flask was added2-methyl-5-t-butylcarbamate aniline 23 (3 g, 13.50 mmol, 1.2 eq),-chloro-3-methyl-4-oxoquinazoline 28 (2.19 g, 11.25 mmol, 1 eq), NaOMe(1.51 g, 15.75 mmol, 1.4 eq), and 2-diphenylbistbutylphosphine (168 mg,5 mol %, 0.561 mmol), and Pd2(dba)3 (309 mg, 3 mol %, 0.336 mmol). Thismixture was refluxed for 8 hours when it was observed there was completeamination of quinazolinone starting material. The reaction mixture wascooled to room temperature and poured into 250 mL EtOAc and then washedwith sodium bicarbonate (2×100 mL), water (2×100 mL), and then brine(2×100 mL). The light brown solid was then purified by flashchromatography on silica (70% EtOAc/Hex) to yield desired product 29 asa light yellow solid (3.6 g, 9.33 mmol, 83% yield). ¹H NMR (300 MHz, D₆DMSO): δ 7.91 (s, 1H), 7.64 (d, 1H, J=2.7 Hz), 7.61 (s, 1H, J=8.8 Hz),7.34 (dd, 1H, J=2.7 Hz, J=8.8 Hz), 7.29 (s, 1H), 7.16 (d, 1H, J=8.2 Hz),7.09 (d, 1H, J=8.2 Hz), 6.47 (s, 1H), 5.70 (s, 1H), 3.59 (s, 3H), 1.64(s, 3H), 1.51 (s, 9H). ¹³C NMR (75 MHz, D₆ DMSO): δ 161.34, 153.57,145.59, 145.58, 141.04, 139.09, 131.84, 129.01, 125.35, 123.96, 123.24,114.31, 112.49, 108.33, 79.70, 34.26, 28.97, 18.14. ESMS: m/z 381.2[MH]⁺

6-(5-Amino-2-methylphenylamino)-3-methylquinazolin-4(3H)-one 30

Tert-butyl4-methyl-3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)phenylcarbamate26 (3.0 g, 7.89 mmol) was added to 50 mL of 10% TFA in DCM. After 1 hr,the reaction mixture was neutralized with 75 mL saturated Na₂CO3,extracted into EtOAc (3×100 mL), dried over MgSO4 and concentrated invacuo to yield the desired product (2.078 g, 7.41 mmol, 94% yield). ¹HNMR (300 MHz, D₆ DMSO): δ 7.91 (s, 1H), 7.71 (d, 1H, J=2.7 Hz), 7.61 (s,1H, J=8.8 Hz), 7.37 (dd, 1H, J=2.7 Hz, J=8.8 Hz), 7.31 (s, 1H), 7.03 (d,1H, J=8.0 Hz), 6.67 (d, 1H, J=2.3 Hz), 6.40 (dd, 1H, J=2.4 Hz, J=8.0Hz), 5.80 (br s, 1H), 3.63 (s, 3H), 2.16 (s, 3H), 1.64 (s, 3H), 1.51 (s,9H). ¹³C NMR (75 MHz, D₆ DMSO): δ 160.73, 146.90, 144.88, 142.13,138.84, 132.85, 131.08, 130.76, 126.96, 125.08, 122.78, 118.25, 115.49,110.06, 34.87, 18.51. ESMS: m/z 281.2 [MH]⁺

4-(4-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-6)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylic acid S-1(0.20 g, 0.73 mmol) was dissolved in DCM (2 mL). To the mixture wasadded HATU (0.41 g, 1.1 mmol) and DIEA (0.38 mL, 2.19 mmol). After 15min of stirring, 4-(4-Amino-phenoxy)-pyridine-2-carboxylic acidmethylamide (LeadGen Labs, LLC; BB-123) (0.18 g, 7.3 mmol) was added.The reaction was stirred overnight at rt. The solvent was evaporated andthe residue was purified by silica gel chromatography (EtOAc/Hx, 0-40%)to yield 57 mg of the desired product, I-6.

4-(4-{[5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonyl]-amino}-phenoxy)-pyridine-2,3-benzimidazolonecarboxylic acid methylamide—(I-12)

5-(4-Fluoro-phenyl)-2-trifluoromethyl-3H-imidazole-4-carbonylic acid S-1(0.05 g, 0.18 mmol) was dissolved in DCM (2.0 mL) and SOCl₂ (2.0 mL).The mixture was refluxed for 2 hr and the solvents were evaporated todryness. The residue was dissolved in DCM and7-(4-aminophenoxy)-1,3-dihydroimidazole[4,5-b]pyridine-2-one (0.08 g,0.33 mmol) and DIEA (0.12 mL, 0.72 mmol) was added. The mixture washeated for overnight and the solvents were evaporated. The residue wasdissolved in DMSO and the product was purified by HPLC to give 8.7 mg ofthe target compound, I-12.

3-(3-methyl-4-oxo-3,4-dihydroquinazolin-6-ylamino)-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamide—(I-13)

In a 20 mL vial equipped with a magnetic stirrer and DCM (3 mL) at 0° C.was added the corresponding imidazole acid S-17 (71 mg, 0.250 mmol, 1eq), EDCI (58.5 mg, 1.5 eq, 0.375 mmol), HOAt (44.25 mg, 1.3 eq, 0.33mmol), DIPEA (175 uL, ˜4 eq, 1.0 mmol). This solution was allowed toreact for 1 hour at 0° C. and then amine (70 mg, 1 Eq, 0.25 mmol) wasadded. The reaction was allowed to react for 24 hours at which time itwas added to 50 mL saturated sodium bicarbonate and extracted into 2×20mL EtOAc. The organic layer was washed with water (20 mL 0.1M HCl) whichextracted some of the color and left a neutral EtOAc extract-1. Theaqueous acidic extract was neutralized with 20 mL saturated NaHCO₃ andback-extracted with 2×10 mL EtOAc to give a second organic extract-2.Both organic extracts were washed with Brine (30 mL), dried overmagnesium sulfate and concentrated to give: Extract-1 totalled 113 mgand extract-2 totalled 30 mg of a dark amber film. Final compound waspurified on a biotage sp1 system utilizing a 25 g silica column and agradient of 2-8% MeOH/CHCl₃ to yield the desired product SFE-0013 (97mg, 0,177 mmol) in 71% yield.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamide—(I-14)

In a 20 mL vial equipped with a magnetic stirrer and DCM (3 mL) at 0° C.was added5-(4-fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylic acidS-17 (71 mg, 0.250 mmol, 1 eq), EDCI (58.5 mg, 1.5 eq, 0.375 mmol), HOAt(44.25 mg, 1.3 eq, 0.33 mmol), DIPEA (175 uL, ˜4 eq, 1.0 mmol). Thissolution was allowed to react for 1 hour at 0° C. and thenN-(2-methyl-5-aminophenyl)-4-(pyridin-3-yl)-pyrimidine-2-amine (AlfaPyridines, AP1C4100) (69 mg, 1 Eq, 0.25 mmol) was added. The reactionwas allowed to react for 24 hours at which time it was added to 50 mLsaturated sodium bicarbonate and extracted and extracted into 2×20 mLEtOAc. The organic layer was washed with water (20 mL 0.1M HCl) whichextracted some of the color and left a neutral EtOAc extract-1. Theaqueous acidic extract was neutralized with 20 mL saturated NaHCO₃ andback-extracted with 2×10 mL EtOAc to give a second organic extract-2.Both organic extracts were washed with Brine (30 mL), dried overmagnesium sulfate and concentrated to give: Final compound was purifiedon a biotage sp1 system utilizing a 25 g silica column and a gradient of2-8% MeOH/CHCl3 to yield the desired product, I-14 (108 mg, 0.199 mmol)in 79% yield.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfonyl-3H-imidazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-15)

Procedure: In a 20 mL vial equipped with a magnetic stirrer and DCM (3mL) at 0° C. was added the corresponding imidazole acid S-17 (71 mg,0.250 mmol, 1 eq), EDCI (58.5 mg, 1.5 eq, 0.375 mmol), HOAt (44.25 mg,1.3 eq, 0.33 mmol), DIPEA (175 uL, ˜4 eq, 1.0 mmol). This solution wasallowed to react for 1 hour at 0° C. and then amine (61 mg, 1 Eq, 0.25mmol) was added. The reaction was allowed to react for 24 hours at whichtime it was added to 50 mL saturated sodium bicarbonate and extractedinto and extracted into 2×20 mL EtOAc. The organic layer was washed withwater (20 mL 0.1M HCl) which extracted some of the color and left aneutral EtOAc extract-1. The aqueous acidic extract was neutralized with20 mL saturated NaHCO₃ and back-extracted with 2×10 mL EtOAc to give asecond organic extract-2. Both organic extracts were washed with Brine(30 mL), dried over magnesium sulfate and concentrated to give an ambersemi-solid. The final compound was purified on a biotage sp1 systemutilizing a 25 g silica column and a gradient of 2-8% MeOH/CHCl3 toyield the desired product I-15 (89 mg, 0.175 mg) 70% yield.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylthio)-1H-imidazole-4-carboxamide—(I-16)

Triethylamine (0.041 mL, 0.29 mmol) was added to a solution of S-15 (60mg, 0.24 mmol), 2-(5-amino-2-methylanilino)-4-(3-pyridyl)pyrimidine (66mg, 0.24 mmol) and HATU (115 mg, 0.29 mmol) in 3 mL DMF. The resultingmixture was stirred at room temperature overnight. Reaction mixture wasthen directly purified on HPLC which afforded 92 mg (76%) of the finalproduct I-16. LC-MS: Rt 1.76 min, 99.8% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfinyl)-1H-imidazole-4-carboxamide—(I-17)

I-17 was prepared from S-16 (50 mg, 0.186 mmol) following proceduredescribed for the synthesis of I-16. After HPLC purification 78 mg (79%)of 1-17 was obtained. LC-MS: Rt 1.51 min, 99% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylthio)-1H-thiazole-4-carboxamide—(I-18)

Diisopropylethylamine (0.484 mL, 2.7 mmol) was added to a solution ofS-25 (150 mg, 0.55 mmol),2-(5-amino-2-methylanilino)-4-(3-pyridyl)pyrimidine (154 mg, 0.55 mmol)and HATU (383 mg, 1.1 mmol) in 6 mL DMF. The resulting mixture wasstirred at room temperature overnight. Reaction mixture was directlysubjected to HPLC purification, which afforded 121 mg (41%) of the finalproduct I-18. LC-MS: Rt 2.21 min, 98% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfinyl)-thiazole-4-carboxamide—(I-19)

Compound I-19 was prepared from S-26 (90 mg, 0.3 mmol) followingprocedure described for the synthesis of 1-18. After HPLC purification29 mg (17%) of 1-19 was obtained. LC-MS: Rt 1.7 min, 100% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonyl)-thiazole-4-carboxamide—(I-20)

Compound I-20 was prepared from S-27 (660 mg, 2.2 mmol) followingprocedure described for the synthesis of I-18. Half of the reactionmixture was purified by HPLC to give 52 mg (4%) of I-20. LC-MS: Rt 1.85min, 97.9% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonylamino)-thiazole-4-carboxamide—(I-21)

Compound I-18 was prepared from S-29 (20 mg, 0.063 mmol) followingprocedure described for the synthesis of I-20. HPLC purification yielded29 mg (80%) of the product I-21. LC-MS: Rt 1.72 min, 99.7% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylthio)-oxazole-4-carboxamide—(I-22)

Compound I-22 was prepared from S-50 (100 mg, 0.4 mmol) followingprocedure described for the synthesis of I-18. After HPLC purification124 mg (61%) of I-22 was obtained. LC-MS: Rt 2.23 min, 98.9% pure byarea.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfinyl)-oxazole-4-carboxamide—(I-23)

Compound I-23 was prepared from S-51 (60 mg, 0.21 mmol) followingprocedure described for the synthesis of I-18. After HPLC purification53 mg (45%) of I-23 was obtained. LC-MS: Rt 1.71 min, 99.8% pure byarea.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonyl)-oxazole-4-carboxamide—(I-24)

Compound I-24 was prepared from S-52 (60 mg, 0.21 mmol) followingprocedure described for the synthesis of I-18. After HPLC purification57 mg (50%) of I-24 was obtained. LC-MS: Rt 1.9 min, 97.3% pure by area.

3-(4-(pyridin-3-yl)-pyrimidinyl-2-yl)amino-4-methyl-analine,5-(4-Fluorophenyl)-2-(methylsulfonyl)-1,2-pyrazole-4-carboxamide—(I-25)

Compound I-25 was prepared from S-60 (156 mg, 0.55 mmol) followingprocedure described for the synthesis of I-18. A portion of the reactionmixture was purified by HPLC to give 17.3 mg (6%) of I-25. LC-MS: Rt1.75 min, 90.1% pure by area.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylthio-1(H)-imidazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-26)

Triethylamine (0.041 mL, 0.29 mmol) was added to a solution of S-15 (60mg, 0.24 mmol), 4-(4-aminophenoxy)pyridine2-carboxilic acid methyamide(58 mg, 0.24 mmol) and HATU (115 mg, 0.29 mmol) in 3 mL DMF. Theresulting mixture was stirred at room temperature overnight. Reactionmixture was then directly purified on HPLC which afforded 66.6 mg (59%)of the final product S-26. LC-MS: Rt 1.94 min, 99.8% pure by area.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfinyl-1(H)-imidazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-27)

Compound I-27 was prepared from S-16 (50 mg, 0.186 mmol) followingprocedure described for the synthesis of I-26. After HPLC purification68 mg (74%) of I-27 was obtained. LC-MS: Rt 1.67 min, 99.9% pure byarea.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylthio-thiazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-28)

Diisopropylethylamine (0.484 mL, 2.7 mmol) was added to a solution ofS-25 (150 mg, 0.55 mmol), 4-(4-aminophenoxy)pyridine2-carboxilic acidmethyamide (135 mg, 0.55 mmol) and HATU (383 mg, 1.1 mmol) in 6 mL DMF.The resulting mixture was stirred at room temperature overnight.Reaction mixture was directly subjected to HPLC purification to afford109 mg (40%) of the product I-28. LC-MS: Rt 2.56 min, 94.7% pure byarea.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfinyl-thiazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-29)

Compound I-29 was prepared from S-26 (90 mg, 0.3 mmol) in the samemanner as 1-28. A portion of the reaction mixture was HPLC purified toprovide 3.6 mg (2%) of the product I-29. LC-MS: Rt 1.94 min, 100% pureby area.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfonyl-thiazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-30)

Compound I-30 was prepared from S-27 (660 mg, 2.2 mmol) in the samemanner as 1-28. Half of the reaction mixture was purified by HPLC togive 48.4 mg (4%) of 11. LC-MS: Rt 2.12 min, 95.4% pure by area.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfonylamino-thiazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-31)

Compound I-31 was prepared from S-29 (20 mg, 0.063 mmol) in the samemanner as I-28. HPLC purification yielded 32 mg (94%) of the productI-31. LC-MS: Rt 1.88 min, 99.4% pure by area.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylthio-oxazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-32)

Compound I-32 was prepared from S-50 (100 mg, 0.4 mmol) followingprocedure described for the synthesis of I-28. After HPLC purification158 mg (84%) of I-32 was obtained. LC-MS: Rt 2.59 min, 99.6% pure byarea.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfinyl-oxazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-33)

Compound I-33 was prepared from S-51 (60 mg, 0.21 mmol) followingprocedure described for the synthesis of I-28. After HPLC purification94 mg (85%) of I-33 was obtained. LC-MS: Rt 1.95 min, 99.8% pure byarea.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfonyl-oxazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-34)

Compound I-34 was prepared from S-52 (60 mg, 0.21 mmol) followingprocedure described for the synthesis of I-28. After HPLC purification61 mg (57%) of I-34 was obtained. LC-MS: Rt 2.16 min, 92.3% pure byarea.

4-(4-{[5-(4-Fluoro-phenyl)-2-methylsulfonyl-1,2-pyrazole-4-carbonyl]-amino}-phenoxy)-pyridine-2-carboxylicacid methylamide—(I-35)

Compound I-35 was prepared from S-60 (156 mg, 0.55 mmol) in the samemanner as I-28. A portion of the reaction mixture was purified by HPLCto give 6.4 mg (2%) of I-35. LC-MS: Rt 2.01 min, 98.2% pure by area.

4-amino-N-(5-(4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamido)-2-methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide(I-36)N-(3-amino-4-methylphenyl)-4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamide.(31)

To a flask containing DCM (23.5 mL) on ice was added4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxylic acid(665 mg, 2.34 mmol) and to this was added portionwise EDC (545 mg, 3.51mmol) then stirred for 10 minutes. To this solution was added HOAt (414mg, 3.04 mmol) followed by DIPEA (1.63 mL, 9.36 mmol). The resultingsolution was allowed to react for 1 hr, at which point4-methylbenzene-1,3-diamine (572 mg, 4.68 mmol) was added and themixture was then warmed to room temperature and stirred for 18 hours. Tothe flask was added saturated sodium bicarbonate (75 mL) then extractedwith DCM (3×30 mL) and EtOAc (3×20 mL). The organic extracts were washedwith brine, combined, dried over Na2SO4, then concentrated under reducedpressure. The crude material was fractionated by SiO2 chromatography(stepwise gradient of DCM to 8% MeOH) and the desired product 30 elutesin 2% MeOH (189 mg, 20% yield). 1HNMR (400 MHz, MeOD/CDCl₃): d 7.72 (m,2H), 7.12 (t, 2H, J=9.2 Hz), 6.94 (d, 1H, J=7.6 Hz), 6.79 (m, 2H), 2.09(s, 3H), 2.02 (s, 3H). 19FNMR (400 MHz, MeOD/CDCl3): d −107.7. MS(ESI):389.0 [M+H]+

4-((1H-benzo[d][1,2,3]triazol-1-yl)oxy)-N-(5-(4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamido)-2-methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide,(32)

To a flask containing MeCN (6.4 mL) and DMF (0.3 mL) on ice was added4-chlorothieno[3,2-d]pyrimidine-7-carboxylic acid (82.9 mg, 0.39 mmol),HOBt (52.2 mg, 0.39 mmol), andn-(3-amino-4-methylphenyl)-4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamide(150 mg, 0.39 mmol). The resulting solution was stirred for 10 minfollowed by the addition of EDC (180 mg, 1.16 mmol) After 48 hrs, TLCindicates that nearly all of the starting material is consumed. To thesolution was added ethyl acetate (10 mL) and water (15 mL) and thesolution was made basic (pH-8) with saturated sodium bicarbonate andextracted with ethyl acetate (3×10 mL). The organic fractions werecombined, washed with brine, dried over Na2SO4, and concentrated underreduced pressure. The crude material was purified by SiO2 chromatography(DCM to 8% MeOH gradient) and the desired product elutes in 3% MeOH (50mg, 22% yield). 1HNMR (400 MHz, CDCl₃/MeOD): d 11.07 (s, 1H), 9.07 (s,1H), 8.89 (s, 1H), 8.69 (s, 1H), 8.14 (s, 1H), 8.01 (d, 1H, J=8.8 Hz),7.75 (m, 2H), 7.54 (m, 1H), 7.44 (m, 2H), 7.33 (d, 1H, J=7.2 Hz), 7.07(d, 1H, J=8.0 Hz), 6.94 (m, 2H), 3.20 (s, 3H), 2.34 (s, 3H). 19FNMR (400MHz, MeOD/CDCl₃): d −110.9. MS(ESI): m/z 683.9 [M+H]+

4-amino-N-(5-(4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamido)-2-methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide(I-36)

Compound4-chloro-N-(5-(4-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-5-carboxamido)-2-methylphenyl)thieno[3,2-d]pyrimidine-7-carboxamide(7.0 mg, 0.01 mmol) was dissolved into 2M ammonia in IPA (1.0 mL, 1.99mmol ammonia). The resulting solution was heated in a sealed vial at 80°C. for 6 hrs. The solvent was removed under reduced pressure and thecrude material was purified by SiO₂ chromatography (DCM to 8% MeOHgradient, desired product elutes in 6% MeOH) (4 mg, 59% yield). 1HNMR(400 MHz, MeOD/CDCl3): d 8.64 (s, 1H), 8.42 (s, 1H), 8.26 (m, 1H), 7.66(m, 2H), 7.56 (m, 2H), 7.51 (s, 1H), 7.41 (m, 2H), 7.07 (d, 1H, J=9.2Hz), 7.01 (t, 2H, J=8.4 Hz), 3.20 (s, 3H), 2.33 (s, 3H). MS(ESI): m/z565.9 [M+H]+.

Preparation of4-(4-(5-(4-fluorophenyl)-1-methyl-2-(methylsulfonyl)-1H-imidazole-4-carboxamido)phenoxy)-N-methylpicolinamide(I-37)

To a chilled solution of4-(4-(5-(4-fluorophenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamido)phenoxy)-N-methylpicolinamide(200 mg, 0.39 mmol) and K2C03 (119 mg, 0.86 mmol) in DMF (3.9 mL), wasadded methyl iodide (29 μL, 67 mg, 0.47 mmol). The reaction mixture wasallowed to warm to room temperature and stirred overnight. TLC indicatescompletion of reaction with a single major product. Solvent was removedunder reduced pressure and the crude material was dissolved in H₂O (20mL) and extracted with EtOAc (3×10 mL). The organic fractions were driedover Na₂SO₄ and concentrated. The crude material was purified by SiO2chromatography (DCM/MeOH from 100% DCM to 94% DCM/6% MeOH, the desiredproduct elutes in 97% DCM/3% MeOH) to yield a white solid (149 mg, 72%yield). 1HNMR (400 MHz, CDCl₃): δ 8.88 (s, 1H), 8.33 (d, 1H, J=6.0 Hz),7.97 (m, 1H), 7.66 (m, 3H), 7.50 (m, 1H), 7.43 (m, 2H), 7.22 (t, 2H,J=8.4 Hz), 7.03 (d, 2H, J=9.2 Hz), 3.79 (s, 3H), 3.50 (s, 3H), 2.98 (d,3H, J=5.2 Hz). MS(ESI): m/z 524.0 [M+H]+

5-(4-Fluorophenyl)-1-methyl-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamide(I-38)

To a chilled solution of5-(4-fluorophenyl)-N-(4-methyl-3-((4-(pyridin-3-yl)pyrimidin-2-yl)amino)phenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamide(200 mg, 0.37 mmol) and K₂CO₃ (112 mg, 0.81 mmol) in DMF (3.7 mL) wasadded methyl iodide (27 uL, 63 mg, 0.44 mmol). The reaction mixture wasallowed to warm to RT and stirred overnight. Solvent was removed underreduced pressure and the crude material was dissolved in H₂O (20 mL) andextracted with EtOAc (3×10 mL). The organic fractions were dried overNa₂SO₄ and concentrated. The crude material was purified by SiO₂chromatography (DCM/MeOH from 100% DCM to 94% DCM/6% MeOH, the desiredproduct elutes in 97% DCM/3% MeOH) to yield a light yellow solid (26 mg,13% yield). 1HNMR (400 MHz, MeOD/CDCl₃): d 9.15 (d, 1H, J=2.4 Hz), 8.56(m, 2H), 8.43 (d, 1H, J=5.2 Hz), 8.31 (s, 1H), 7.60 (s, 1H), 7.47 (m,2H), 7.27 (m, 3H), 7.19 (t, 3H, J=8.4 Hz), 3.77 (s, 3H), 3.46 (s, 3H),2.28 (s, 3H). MS(ESI): m/z 558.0 [M+H]+

4-(4-Fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-(trifluoromethyl)oxazole-5-carboxamide.(I-39)

To a flask containing DCM (3 mL) on ice was added4-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-5-carboxylic acid (S-41)(76 mg, 0.28 mmol), EDC (47.2 mg, 0.3 mmol), HOAt (48.9 mg, 0.36 mmol),and DIPEA (0.193 mL, 1.10 mmol). The resulting solution was stirred for1 hr followed by the addition of6-((5-amino-2-methylphenyl)amino)-3-methylquinazolin-4(3H)-one (HGM-7)(77.4 mg, 0.28 mmol). The reaction was stirred for 18 hours at ambienttemperature at which point the solution was made basic by the additionof saturated sodium bicarbonate (10 mL) which was then extracted withDCM (2×10 mL) and EtOAc (2×10 mL). The organic extracts were washed withbrine, combined, dried over Na₂SO₄, and concentrated. The crude materialwas fractionated by SiO₂ chromatography (stepwise gradient of DCM to 90%DCM/10% MeOH, desired product elutes in 5% MeOH)(101 mg, 67% yield).1HNMR (400 MHz, CDCl₃): d 8.9 (s, 1H), 7.82 (m, 2H), 7.80 (s, 1H), 7.55(d, 1H, J=2.4 Hz), 7.50 (s, 1H), 7.41 (d, 1H, J=8.4 hz), 7.27 (m, 2H),7.03 (d, 1H, J=8.4 Hz), 6.95 (t, 2H, J=8.8 Hz), 3.44 (s, 3H), 2.12 (s,3H).

5-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-(trifluoromethyl)oxazole-4-carboxamide.(I-40)

To a flask containing DCM (3 mL) at 0° C. was added5-(4-fluorophenyl)-2-(trifluoromethyl)oxazole-4-carboxylic acid (S-68)(17 mg, 0.06 mmol), EDC (10.6 mg, 0.07 mmol), HOAt (10.9 mg, 0.08 mmol),and DIPEA (0.043 mL, 0.25 mmol). The resulting solution was allowed toreact at 0° C. for 1 hr, at which point6-((5-amino-2-methylphenyl)amino)-3-methylquinazolin-4(3H)-one(HGM-7)(17.3 mg, 0.06 mmol) was added. The reaction was allowed to reactfor 18 hours at ambient temperature at which point saturated sodiumbicarbonate (5 mL) was added and extracted with DCM (2×10 mL) then EtOAc(2×10 mL). The organic extracts were washed with brine, combined, driedover Na₂SO₄, and concentrated. The crude material was fractionated bySiO₂ chromatography (stepwise gradient of DCM to 10% MeOH, the desiredproduct elutes in 6% MeOH) (9 mg, 27% yield). 1HNMR (400 MHz,CDCl₃/MeOD): d 8.29 (m, 2H), 7.98 (s, 1H), 7.62 (d, 1H, J=2.4 Hz), 7.57(m, 2H), 7.51 (s, 1H), 7.42 (dd, 1H, J=2.8, 9.2 Hz), 7.39 (dd, 1H,J=2.4, 8.4 Hz), 7.24 (d, 1H, J=8.0 Hz), 7.19 (t, 2H, J=8.4 Hz), 3.58 (s,3H), 2.24 (s, 3H).

5-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-2-(methylsulfonyl)oxazole-4-carboxamide,I-41

6-(5-amino-2-methyl-phenylamino)-3-methyl-3H-quinazolin-4-one HCl salt(110 mg, 0.28 mmol) was dissolved in DMF (5 mL) with5-(4-fluoro-phenyl)-2-methanesulfonyl-oxazole-4-carboxylic acid (142 mg,0.5 mmol) at RT. HATU (150 mg, 0.39 mmol) was added, followed by DIEA(0.5 mL, 2.87 mmol). The mixture was stirred for 20 h. Water (20 mL) wasadded and the mixture was extracted with EtOAc (30 mL). The organiclayer was concentrated and the residue was dissolved in minimum amountof DMSO and purified by prep HPLC. 82 mg of title compound was obtainedafter lyophilization of pure fractions. LC-MS [M+H] 548.1.

2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-1H-imidazole-4-carboxamide,I-42

6-(5-Amino-2-methyl-phenylamino)-3-methyl-3H-quinazolin-4-one HCl salt(662 mg, 1.7 mmol) was dissolved in DMF (15 mL) with2-(2-dimethylamino-ethylthio)-5-(4-fluoro-phenyl)-1H-imidazole-4-carboxylicacid (750 mg, 2.55 mmol) at RT. HATU (950 mg, 2.5 mmol) was added,followed by DIEA (1.5 mL, 8.6 mmol). The mixture was stirred for 16 h.Water (100 mL) was added and the mixture was extracted with EtOAc (300mL). The organic layer was concentrated and the residue was dissolved inminimum amount of DMSO and purified by prep HPLC. 700 mg of titlecompound was obtained after lyophilization of pure fractions. LC-MS[M+H] 572.2.

2-((2-(dimethylamino)ethyl)sulfinyl)-5-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-1H-imidazole-4-carboxamide,I-43

2-(2-Dimethylamino-ethylthio)-5-(4-fluoro-phenyl)-1H-imidazole-4-carboxylicacid[4-methyl-3-(3-methyl-4-oxo-3,4-dihydro-quinazolin-6-ylamino)-phenyl]-amide(150 mg, 0.22 mmol, TFA salt) was dissolved in a mixed solvent ofDCM/CH₃CN (1.5 mL each) at RT. Oxone (80 mg, 0.13 mmol) was added,followed by MeOH (10 drops). The mixture was heated to 60° C. for 40mins. It was then cooled and concentrated. The residue was purified byprep HPLC to afford 127 mg of the title compound. LC-MS [M+H] 588.2.

2-((2-(dimethylamino)ethyl)sulfonyl)-5-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-1H-imidazole-4-carboxamide,I-44

2-(2-Dimethylamino-ethylthio)-5-(4-fluoro-phenyl)-1H-imidazole-4-carboxylicacid[4-methyl-3-(3-methyl-4-oxo-3,4-dihydro-quinazolin-6-ylamino)-phenyl]-amide(110 mg, 0.26 mmol, TFA salt) was dissolved in a mixed solvent ofDCM/CH₃CN (1.5 mL each) at RT. Oxone (118 mg, 0.26 mmol) was added,followed by a mixture of MeOH/water (2/1 6 drops). The mixture washeated to 60° C. for 3 h. It was then cooled and concentrated. Theresidue was purified by prep HPLC to afford 60 mg of the title compound.LC-MS [M+H] 604.2.

3-bromo-1-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-1H-pyrazole-5-carboxamide,I-45

6-(5-Amino-2-methyl-phenylamino)-3-methyl-3H-quinazolin-4-one HCl salt(30 mg, 0.08 mmol) was dissolved in DMF (5 mL) with5-bromo-2-(4-fluoro-phenyl)-2H-pyrazole-3-carboxylic acid (45 mg, 0.16mmol) at RT. HATU (60 mg, 0.16 mmol) was added, followed by DIEA (0.14mL, 0.8 mmol). The mixture was stirred for 20 h. Water (10 mL) was addedand the mixture was extracted with EtOAc (20 mL). The organic layer wasconcentrated and the residue was dissolved in minimum amount of DMSO andpurified by prep HPLC. 30 mg of title compound was obtained afterlyophilization of pure fractions. LC-MS [M+H] 549.0.

1-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-(methylthio)-1H-pyrazole-5-carboxamide,I-46

6-(5-amino-2-methyl-phenylamino)-3-methyl-3H-quinazolin-4-one HCl salt(110 mg, 0.28 mmol) was dissolved in DMF (5 mL) with2-(4-fluoro-phenyl)-5-methylthio-2H-pyrazole-3-carboxylic acid (125 mg,0.5 mmol) at RT. HATU (150 mg, 0.39 mmol) was added, followed by DIEA(0.5 mL, 2.87 mmol). The mixture was stirred for 20 h. Water (20 mL) wasadded and the mixture was extracted with EtOAc (30 mL). The organiclayer was concentrated and the residue was dissolved in minimum amountof DMSO and purified by prep HPLC. 80 mg of title compound was obtainedafter lyophilization of pure fractions. LC-MS [M+H] 514.2.

1-(4-fluorophenyl)-N-(4-methyl-3-((3-methyl-4-oxo-3,4-dihydroquinazolin-6-yl)amino)phenyl)-3-(methylsulfonyl)-1H-pyrazole-5-carboxamide,I-47

6-(5-Amino-2-methyl-phenylamino)-3-methyl-3H-quinazolin-4-one HCl salt(138 mg, 0.49 mmol) was dissolved in DMF (2 mL) with2-(4-fluoro-phenyl)-5-methanesulfonyl-2H-pyrazole-3-carboxylic acid (140mg, 0.49 mmol) at RT. HATU (186 mg, 0.54 mmol) was added, followed byDIEA (0.21 mL, 1.2 mmol). The mixture was stirred for 20 h. Water (10mL) was added and the mixture was extracted with EtOAc (20 mL). Theorganic layer was concentrated and the residue was dissolved in minimumamount of DMSO and purified by prep HPLC. 55 mg of title compound wasobtained after lyophilization of pure fractions. LC-MS [M+H] 547.2.

5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-2-(methylsulfonyl)-1H-imidazole-4-carboxamide,I-48

3-Imidazo[1,2-b]pyridazin-3-ylethynyl-4-methyl-phenylamine TFA salt (200mg, 0.55 mmol) was dissolved in DMF (5 mL) with5-(4-fluoro-phenyl)-2-methanesulfonyl-1H-imidazole-4-carboxylic acid(204 mg, 0.72 mmol) at RT. HATU (300 mg, 0.79 mmol) was added, followedby DIEA (0.6 mL, 3.44 mmol). The mixture was stirred for 20 h. Water (10mL) was added and the mixture was extracted with EtOAc (30 mL). Theorganic layer was concentrated and the residue was dissolved in minimumamount of DMSO and purified by prep HPLC. 140 mg of title compound wasobtained after lyophilization of pure fractions. LC-MS [M+H] 515.2.

2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-1H-imidazole-4-carboxamide,I-49

3-Imidazo[1,2-b]pyridazin-3-ylethynyl-4-methyl-phenylamine TFA salt (450mg, 1.24 mmol) was dissolved in DMF (10 mL) with2-(2-Dimethylamino-ethylthio)-5-(4-fluoro-phenyl)-1H-imidazole-4-carboxylicacid (575 mg, 1.86 mmol) at RT. HATU (700 mg, 1.86 mmol) was added,followed by DIEA (1.3 mL, 7.44 mmol). The mixture was stirred for 20 h.Water (80 mL) was added and the mixture was extracted with EtOAc (300mL). The organic layer was concentrated and the residue was dissolved inminimum amount of DMSO and purified by prep HPLC. 540 mg of titlecompound was obtained after lyophilization of pure fractions. LC-MS[M+H] 540.3.

2-((2-(dimethylamino)ethylsulfinyl)-5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-1H-imidazole-4-carboxamide,I-50

2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-1H-imidazole-4-carboxamide(200 mg, 0.31 mmol, TFA salt) was dissolved in a mixed solvent ofDCM/CH₃CN (1.5 mL each) at RT. Oxone (95 mg, 0.16 mmol) was added,followed by MeOH (10 drops). The mixture was heated to 60° C. for 40mins. It was then cooled and concentrated. The residue was purified byprep HPLC to afford 121 mg of the title compound. LC-MS [M+H] 556.0.

2-((2-(dimethylamino)ethylsulfonyl)-5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-1H-imidazole-4-carboxamide,I-51

2-((2-(dimethylamino)ethyl)thio)-5-(4-fluorophenyl)-N-(3-(imidazo[1,2-b]pyridazin-3-ylethynyl)-4-methylphenyl)-1H-imidazole-4-carboxamide(200 mg, 0.31 mmol, TFA salt) was dissolved in a mixed solvent ofDCM/CH₃CN (2.0 mL each) at RT. Oxone (380 mg, 0.62 mmol) was added,followed by a mixture of MeOH/water (2/1, 6 drops). The mixture washeated to 60° C. for 3 h. It was then cooled and concentrated. Theresidue was purified by prep HPLC to afford 128 mg of the titlecompound. LC-MS [M+H] 572.2.

New Binding Mode Hypothesis Points to Uniqueness:

The unique properties of this scaffold geometry of compounds of Formula(I) are believed to result from its novel mode of binding to the DFG-outconformation adopted by many kinases. In contrast to the previouslyproposed binding mode (Dietrich, 2010), FIG. 8A, we herein describe asignificantly different binding mode hypothesis, FIG. 8-B.

FIG. 8 illustrates differences between the previously-described B-Rafbinding mode hypothesis for KIN-035 (Dietrich et. el., 2010), panel-A,and the new binding mode described herein, panel-B which are 1) thelower selectivity site is occupied by the 4-F-phenyl substituent ratherthan the 2-trifluoromethyl substituent on the imidazole ring, 2) theGlu⁵⁰⁰ side-chain is not required to move and the carboxamide linkagecan bridge, in the typical fashion, between the Glu⁵⁰⁰ side chain andthe NH of Asp⁵⁹³. 3) a different orientation of the quinazolinone, thusallows both hydrogen bonding to Cys⁵³² and Thr⁵²⁸. As indicated inChart-1, KIN-035, was a potent inhibitor of CSF1R but the other twoinhibitors, which varied only slightly in structure, were not. A deeperlook into the reported data (Dietrich, 2010) suggested that the additi9.

In particular, addition of a 4-fluoro-substituent on the imidazolephenyl ring, ie. KIN-032 vs KIN-035, improved potency 70-fold.Conversely, replacement of the 4-fluoro substituent on the imidazolephenyl ring with larger R³ and R⁴ groups dramatically decreases potencyfor CSF1R.

FIG. 9 provides a summary of SAR data for imidazole quinazolinoneinhibitors where there is a dramatic decrease in CSF1 inhibition when R³and R⁴ substituents increase in size.

The recent release of the first co-crystal structure of CSF1R with aType-II inhibitor (3LCO.pdb) provided an opportunity to see if the newbinding mode hypothesis described herein accounted for theseobservations. Modeling of KIN-035 into this crystal structure did indeedindicate an unusually tight fit of the 4-fluoro substituent into anindentation at the bottom of the lower selectivity site, FIG. 9C.

FIG. 10 shows models of KIN-035 in A) B-Raf, B) p38-α, and C) CSF1R anda new inhibitor, I-6, modeled into D) KDR(VEGFR2), E) FLT3, and F)c-Met. DFG-loops are shown in mauve, conserved lysines and glutamateresidues are shown in blue and red, and lower selectivity site residuesshown in green CPK form. The arrow illustrates where 4-fluorosubstituent fits in indentation.

A similar analysis of other co-crystal structures with Type-IIinhibitors, FIG. 10, revealed that an indentation at the bottom of thelower selectivity site, which is typically constructed from 3 to 4branched aliphatic amino acid side chains shown in green, is a highlyconserved feature. It was also observed that variations in the size ofthis indentation existed between kinase structures and that the uniquegeometry of the ortho-aryl-imidazole carboxamide scaffold orients the4-fluoro-aryl substituent ideally to interact with this indentation. Forcomparison, many c-Met inhibitors contain a 4-F-phenyl substitutedphenyl ring that occupies the lower selectivity site, see co-crystalstructure of BMS-777607 in 3F82.pdb, (Raeppel S., Bioorg. Med. Chem.Letts. (2009), 19:1323-1328; Schroeder G M., J. Med. Chem. (2009)52(5):1251-1254). However, detailed analysis reveals that the fluorineatom in these compounds are not positioned to interact with theindentation at the bottom of the lower selectivity site of c-Met, FIG.11.

In spite of structural similarity of these inhibitors, c-Met inhibitorsA and B do not penetrate into the indentation at bottom of lowerselectivity site, see 3F82.pdb, where as inhibitor 12 can reach thisindentation due to its different scaffold geometry (FIG. 11).Conversely, the carbonyl H-bond interactions with the gatekeeper regionlysine side-chain exhibited by inhibitors A) and B) is absent ininhibitor I-6 unless mediated by a bridging water.

Implications from Biological Activities:

Broad Applicability of Ortho-Aryl-Imidazole Carboxamide Scaffold:

A total of 15 new inhibitors were designed, prepared, and evaluatedagainst a panel of 29 selected kinases in order to explore theapplicability, SAR trends, and the tunability of this scaffold thatcould be imposed but this unique geometry, Table 21. Inhibitors I-1 to1-12 all retained the 2-CF₃-5-(4-F-phenyl) imidazole-4-carboxamidescaffold previously described (Deitrich, 2010), but explored differentHinge-Gatekeeper Motifs (HGM). Predictions from the molecular modelinganalysis of KDR, FLT-3, and c-Met, FIG. 10, suggested inhibitors I-6(Sorafenib HGM) and I-12, should inhibit a broader set of kinases thanpreviously described (Dietrich, 2010).

Inhibitor I-13 was designed to be a direct analog of previously reportedKIN-35 (Dietrich et. al., 2010) which incorporates the significantlymore polar 2-CH₃SO₂-5-(4-F-phenyl) imidazole-4-carboxamide scaffold. Dueto the significant shift in polarity compared to the 2-CF3-substituent,a major objective of inhibitor I-13 was to test the proposed new bindingmode hypothesis. Inhibitor I-14 was designed to evaluate how attachmentof this new polar scaffold to the pyridyl-pyrimidine containing HGM ofImatinib would compare to the known profile of activities for this drug.Inhibitor I-15 was designed to evaluate how attachment of this polarscaffold would compare with that seen for Inhibitor I-6.

Because molecular modeling studies of the new binding mode, FIG. 8,suggests a tight fit of the ortho-4-fluorophenyl-5-membered heterarylcarboxamide scaffold into the lower selectivity site, it was suspectedthat the HGM variations could display unique SAR and unexpectedselectivity profiles due to severely limited active site dynamics.Molecular modeling studies also suggested that replacement of the2-trifluoromethyl group on the imidazole scaffold by the methylsulfonylmoiety would be a good test of the proposed new binding mode hypothesis.The results from this limited kinase profiling are summarized inTable-21.

TABLE 21 Comparison of % inhibition profiles of KIN035 at 1 μM andInhibitors 1-13 at 5 μM. Kinases Profiled Inhib- FMS itor DCAM- AUROR-MEK- DDR- EPH- FLT- FLT- FLT- (CS- IG- Structure # KL2 JNK2 MAPK1 P38αP13K_(α) A 1 ABL1 BTK 2 EGFR A2 1 3 4 F1R) F1R KDR KIT Ambit Biosciences% Inhibition at 1 μM inhibitor concentration without prolongedpreincubation

KIN- 035 X X X X X X X X X X X X X X X X Nanosyn Inc. % Inhibition at 10μM inhibitor concenration without prolonged preincubation

I-1  X

I-2 

I-3  X X X X X X X X X

I-4  X X X X X

I-5  X X X X

I-6  X X X X X X X X X X X X X X X X X X X X X X X X X X X X

I-7  X X

I-8  X X X X X X X X X X X X

I-9  X X X X X X X X

I-10 X X X X X X

I-11 X X X X X X

I-12 X X X X X X X X X X X X X X X X X X X X X X X X

I-13 X X X X X X X X Inhib- Kinases Profiled itorBRAF- >25% >50% >75% >100% Structure # MET PDGFRα PDGFRβ RET RON TIE2TYK2 BRAF V599E CRAF Inhib. Inhib. Inhib. Inhib. Ambit Biosciences %Inhibition at 1 μM inhibitor concentration without prolongedpreincubation

KIN- 035 X X X X X X X X X X X X X X X X X X X X X X X X X X X X X X XNanosyn Inc. % Inhibition at 10 μM inhibitor concenration withoutprolonged preincubation

I-1  X X X X X X X X X X X

I-2  X X X X X X X X X X

I-3  X X X X X X X X X X X X X X X X

I-4  X X X X X X X X X X X X X X

I-5  X X X X X X X X X X X X

I-6  X X X X X X X X X X X X X X X X X X X X X X X

I-7  X X X X X X X X X X

I-8  X X X X X X X X X X X X X X X X

I-9  X X X X X X X X X X X X X X X X X X X X

I-10 X X X X X X X X X X X X X

I-11 X X X X X X X X X X X X X X

I-12 X X X X X X X X X X X X X X X X X X X X X X X

I-13 X X X X X X X X X X X X X X X X X X X X

As anticipated, inhibitor I-6, which incorporated the HGM of Sorafenib,displayed a significantly expanded selectivity profile compared toKIN-035 (Deitrich et. al., 2010), which, with some exceptions, resembledthat of Sorafenib itself. Compared to the activity profile reported forSorafenib (DDR2, FLTs(1,3,4), FMS, KDR, KIT, PDGFR-α, PDGFR-0, and RETwith some ABL1, B-Raf, C-Raf, and TIE2, but not c-MET or RON activities(Karaman, 2008; Kumar, 2009), inhibitor I-6 most potently inhibitedDDR2, FLTs(1,3,4), FMS, KDR, KIT, PDGFR-α, PDGFR-β, and TIE2 with someABL1, B-Raf, C-Raf, and RET with the addition of c-MET and RONactivities. While these modeling studies predicted that KDR and FLT-3,as well as many other kinases, should be inhibited, it was not expectedthat c-Met inhibition be observed owning to the lack of an appropriategroup to interact with Lys 1110, see 3F82.pdb (Schroeder G. M., J. Med.Chem. (2009) 52(5):1251-1254).

The related inhibitor I-12 also inhibited many of the same kinases asinhibitor I-6 with the best inhibition seen for DDR2, FLT-3, FMS, andTIE2. More modest activity for Aurora-A, FLT-1/4, KDR, KIT, PDGFRs wasaccompanied by better activity for B-Raf mutant and wild-type forms,Modest inhibition of MEK-1, BTK, c-Met, RET, and RON was also observed.These data support our new binding hypothesis and demonstrates that thisortho-(4-F-phenyl)-5-membered heteroaryl carboxamide scaffold has broadapplicability. Moreover, the observed inhibition of MEK-1 with 1-12 andTYK2 with I-6 represents additional activities against kinases for whichno type-II inhibitors have been reported to date. These data suggeststhat this unique scaffold geometry may better stabilize the DFG-outconformation of kinases and could be useful to probe kinases for thisconformational change. To further evaluate the scope of inhibition, I-6was evaluated against a larger panel of 230 kinases, Table-22. Thesedata better illustrate the potential applicability of the scaffoldgeometry to inhibitor design.

TABLE 22 Kinases inhibited at least 50% by I-6 when evaluated at 5 uMagainst a panel of 230 kinases. Kinases are listed in alphabeticalorder. ABL1 ABL-T315I ARG AXL BLK BRAF CRAF DDR2 EPH-A1 EPH-A2 EPH-A3EPH-A4 EPH-A5 EPH-A8 EPH-B1 EPH-B2 EPH-B4 FLT1 FLT3 FLT4 FGR FER FMS HCKHIPK3 HIPK4 JAK2 KDR KIT LCK LOK LYNA LYNB MAP4K2 MAP4K5 MEK2 MER METMKNK1 MNK2 MUSK P38-α PAK1 PDGFR-α PDGFR-β PRKD3 PTK5 RET RON SIK SPHK1SPHK2 SRC SRMS TIE2 TRKA TRKB TRKC TYK2 TYRO3 YES

The reverse amide series of inhibitors I-1 to I5 was prepared becausecompounds with these HGMs had displayed good activity against FMS (LyneP. D., Bioorg. Med/Chem. Letts. (2009), 19(3): 1026-1029). Surprisingly,of these five compounds only I-3 demonstrated significant inhibition ofFMS while I-1 and I-2 showed no significant activity against for any ofthe kinases tested. This poor activity is thought to result from therigid anchoring of the trisubstituted imidazole scaffold described inthe new binding mode hypothesis described herein. In addition, severalnoteworthy combinations of activities were observed. Noteworthy amongthese are:

-   -   1) Inhibitor 6: Inhibition, albeit modest, of TYK2.    -   2) Inhibitor 11: High selectivity for p38α.    -   3) Inhibitor 11: Observation of PI3K-α inhibition.    -   4) Inhibitor 12: Inhibition, albeit modest, of MEK1, a kinase        for which no Type-II inhibitors have been reported.    -   5) Inhibitor 13: Observation of PI3K-α inhibition with the        2-methylsulfone analog of KIN-035.    -   6) Inhibitor 13: The unique combination of p38-α, B-Raf, C-Raf,        and PI3K activities.

Considerations Specific to B-Raf(V600) Activity:

As previously stated, type-II kinase inhibitors bind to a minorconformation some kinases exhibit in which a DFG-loop vacates alipophilic pocket immediately adjacent to the ATP binding site. Thisloop movement is illustrated in FIG. 2.

In contrast to the previously reported binding mode utilizing 3C4C.pdb,FIG. 12A, our molecular modeling studies were based on the co-crystalstructures (1UWJ.pdb) of B-Raf (V600E) with Sorafenib and the morerecent co-crystal structure (4G9R.pdb) of B-Raf(V600E) with thequinazolinone inhibitor AZ628. These studies provided an alternativemode of binding depicted in FIG. 12B. Detailed examination of thepotential interactions available to this scaffold that are unique to theV600E mutant structures compared to structures of the wild type kinase,suggested a strategy for the development of selective inhibitors ofB-Raf(V600E). A key difference between wild type B-Raf and the valine togluatate (V600E) mutant in various co-crystal structures is thepositioning of the adjacent Lys601 residue. In the wild type kinasestructure 4JGV.pdb, this residue is solvent exposed and on the surfaceto the protein, FIG. 13A, whereas in the two B-RafV600E structures(1UWJ.pdb and 4G9R.pdb) this residue is flipped in toward the inhibitor,FIG. 13B.

To test both our binding mode hypothesis and to see if these structuraldifferences could be taken exploited to impart selectivity and potencyimprovements, I-13, in which the lipophilic 2-trifluoromethy substituenton imidazole ring of inhibitor KIN-035, was replaced by the polar2-methylsulfonyl substitutent. In a side by side IC₅₀ comparison using aLanthaScreen assay, a 10 fold increase in potency (IC₅₀=6.6 nM for I-13vs. 67 nM with KIN-0035) was observe. In addition, I-13 wasapproximately 80 fold more potent against the mutant verses wild typeB-Raf (IC₅₀˜500 nM). Interestingly, it was observed that the IC₅₀ valuefor I-13 appeared to decrease over a several hour preincubation period.To address this issue, real time kinetic experiments were performed.These kinetic data, FIG. 13, are consistent with either extremely tightbinding or irreversible inhibition.

The kinetic data with I-13 are consistent with the reported longdisassociation half life for AZ-628 with which it shares thequinazolinone Hinge Interacting motif (Hatzivassiliou G. et al, Nature(2010) 464: 431-435. It is feasible that, with a minor movement of theLys⁶01 sidechain, FIG. 14A, the terminal amino group can be positionedfor favorable electrostatic interactions with the 2-methylsulfonyl groupof I-13. Furthermore, it is mechanistically feasible for the side-chainamino group of this residue to attack the imidazole ring displacing themethylsulfonyl group in order to form a covalent adduct, FIG. 14B. Thenucleophilic aromatic substitution of methyl sulfonyl groups adjacent topyridine or pyrimidine nitrogens is well known and the reaction of2-methylsulfonyl-benzimidazole with amines has been reported, albeitwith strong base, at elevated temperatures (140 deg. C.), and in theabsence of a solvent. (Ping L. et al, Tetrahedron Lett. (2008) 49(12):1910-1914). If such a displacement were to take place, it would likelyrequire a specific geometry, prolonged contact time, and catalyticassistance from neighboring groups at the active site. FIG. 13illustrates how such a convergence might be achieved with B-Raf (V600E).Regardless of mechanism, exploitation of these differences between wildtype and V600E B-Raf could lead to inhibitors for the treatment ofcancers with improved potency and selectivity.

Because c-Raf, PI3K-α and MEK1 are kinases reported to enable the escapeof B-Raf mutant addicted cancers from B-Raf(V600E) inhibition, discoveryof a B-Raf inhibitor with these additional activities represents a veryattractive and unique opportunity for anti-cancer therapy. Takentogether, these data illustrate that, even within a limited number ofanalogs, unexpected and unique activities can be obtained using thisnovel scaffold geometry.

To further test the breadth of applicability and ability to achieveselectivity with inhibitors that share this scaffold geometry, I-11 andI-12 were evaluated against a larger panel of 220 high interest kinases,Table-23.

TABLE 23 Summary of profiling data for I-11, I-12, I-14 and I-15 assayedin duplicate against a panel of 220 kinases. Kinases listed in tablewere inhibited better than 50% at the profiling concentration of 5 μM.

Inhibitor 11 Kinase Cmpd Conc. (μM) Target 5 KIT-V560G 5 P38-α

Inhibitor 12 Cmpd Conc. (μM) Kinase Target 5 ABL-M351T 5 ABL-Q252H 5ABL-H396P 5 ARG 5 AURORA-A 5 AURORA-C 5 BRK 5 BLK 5 CDK2 5 DDR2 5 EPH-A45 EPH-B2 5 EPH-B4 5 FER 5 FGR 5 FLT-1 5 FLT-3 5 FMS 5 HIPK4 5 KDR 5 KIT5 KIT-V560G 5 LCK 5 LOK 5 LYNA 5 LYNB 5 MAP4K2 5 MER 5 MET 5 MKNK1 5MNK2 5 MUSK 5 P38-α 5 PDGFR-α 5 PDGFR-α-V561D 5 PTK5 5 RON 5 SRMS 5 SRC5 TIE2 5 TNK1 5 TRKA 5 TRKB 5 TRKC 5 TYRO3

Inhibitor 14 Cmpd Conc. (μM) Kinase Target 5 ABL-Q252H 5 ABL-M351T 5 ARG5 BRAF-V599E 5 CRAF 5 DDR2 5 FMS 5 KIT 5 KIT-V560G 5 PI3-KINASE-α 5 TNK15 PI3-KINASE-δ 5 P38-α 5 PDGFR-α-V561D 5 PDGFR-α 5 PDGFR-β

Inhibitor 15 Cmpd Conc. (μM) Kinase Target 5 ABL-Q252H 5 ABL-M351T 5ABL-H396P 5 BLK 5 BRK 5 DDR2 5 FMS 5 EPH-A4 5 EPH-B2 5 MUSK 5 HIPK4 5LOK 5 MER 5 MNK2 5 MUSK 5 SRMS 5 TIE2 5 TRKA 5 TRKB 5 TRKC 5 TYRO3

While I-11 remained extremely selective for p38-α, I-12 displayedgreater that 50% inhibition for 45 of the panel kinases and 72 kinasesat >25% inhibition. These data illustrate that, by modifying the HGM,both a wide range of kinases can be inhibited and high degrees ofselectivity can be obtained. Inhibitors, I-14 and I-15 were also testedagainst this panel, Table-22. These two inhibitors illustrate thatattractive profiles can be obtained using a wider range of HGMs.

Because the inhibition profile of I-14 included members of both the MAPKand PI3K pathways, IC₅₀ determinations were performed on the mostsignificantly inhibited kinases with the inclusion of all the PI3Kisoforms. These data, Table-24 which were intended to explore bothpotency and mechanism of inhibition, revealed the most significant timedependence with all the PI3K isoforms. The observation of potency andisoform selectivity with time-dependency for PI3K-γ appears to beunprecedented in the literature. In addition, these activities combinedwith inhibition of p38-α, Raf-1, DDR and CSF-1R, would be complimentaryand potentially synergistic activities for both the treatment of certaininflammation based conditions and cancers.

TABLE 24 Determination of time dependence and IC50 values for I-14 forselected kinases. Due to the low ATP concentration used (10 uM), theeffects of preincubation may be attenuated for some kinases. CompoundCompound Time Inhibitor I-14 IC50* (nM): IC50* (nM): DependencePreincubation 15 Min 60 Min Lipid Kinase I-14 I-14 Observed? PI3K-γ126.60 57.82 Yes PI3K-α 898.90 386.60 Yes PI3K-δ 3177.00 1121.00 YesPI3K-β 12620.00 2908.00 Yes Protein Kinases RAF1 96.55 81.35 YesP38a/MAPK14 149.70 150.10 No BRAF (V599E) 194.40 194.00 No DDR2 222.30221.10 No FMS 219.90 239.60 No TNK1 210.00 241.00 No BRAF 388.30 323.10Yes PDGFR-α 177.70 342.00 No c-Kit 772.10 986.90 No P38b/MAPK11 1018.001084.00 No ROS/ROS1 1248.00 1284.00 No ARAF 1394.00 1296.00 Yes PDGFR-β2014.00 2512.00 No Aurora A 2738.00 2633.00 No

Based on the interesting activity data for I-14 land I-15, a smalllibrary of 20 analogs was prepared, ie. 10 new scaffolds coupled to theImatinib and Sorafenib HGMs (HGM-1 and HGM-17), to explore the effectsof scaffold alterations on activity profiles, Table 25. A summary ofthese activity profiles compared to those of I-14 and I-15 is presentedin Table 26 and Table 27.

TABLE 25 Construction of the Expanded Library of 20 New Inhibitors basedon inhibitors I-14 and I-15.                   Scaffold #                  Scaffold Structure

                  HGM-1 Inhibitor Structure S-15

I-16

S-16

I-17

S-17

I-14

S-25

I-18

S-26

I-19

S-27

I-20

S-29

I-21

S-50

I-22

S-51

I-23

S-52

I-24

S-60

I-25

                  Scaffold #

                HGM-35 Inhibitor Structure S-15 I-26

S-16 I-27

S-17 I-15

S-25 I-28

S-26 I-29

S-27 I-30

S-29 I-31

S-50 I-32

S-51 I-33

S-52 I-34

S-60 I-35

TABLE 26 Kinase Inhibition Profiles of the 20 new compounds defined inTable 25 against a panel of 230 kinaes when assayed at 5 uM. Theinhibiton profiles for inhibitor I-15 is included for comparison.Scafffold # 15 16 17 25 26 27 29 50 51 52 60 HGM# 1 1 1 1 1 1 1 1 1 1 1Cpd # I-16 I-17 I-14 I-18 I-19 I-20 I-21 I-22 I-23 I-24 I-25 ABL1 ARGARG ARG ABL1 ABL1 ARG ABL1 ABL1 ABL1 ABL1 ARG AURORA-A CRAF CRAF ARG ARGCRAF ARG ARG ARG ARG CRAF AURORA-B DDR2 DDR2 ARK5 CLK4 DDR2 CRAF ARK5BTK ARK5 DDR2 AXL FMS EPH-A5 AXL CRAF FMS DDR2 AXL CRAF CRAF EPH-A2 CLK4KIT EPH-A8 CDK2-Cy DDR2 KIT EPH-A2 CDK2-Cy DDR2 DDR2 EPH-A4 CRAF P38-αEPH-B2 CK2 EPH-A2 P38-α EPH-A3 CK2 EPH-A2 EPH-A2 EPH-A5 DDR2 PDGFR-α KITCLK4 EPH-A4 P38-β EPH-A4 CLK4 EPH-A4 EPH-A4 EPH-A8 EPH-A2 PDGFR-βPDGFR-α CRAF EPH-A5 PDGFR-α EPH-A5 CRAF EPH-A5 EPH-A5 EPH-B1 EPH-A4PI3K-α PDGFR-β DDR2 EPH-A8 ROS EPH-A8 DDR2 EPH-A8 EPH-A8 EPH-B2 EPH-A5TNK1 EPH-A1 EPH-B1 TNK1 EPH-B1 EPH-A1 EPH-B1 EPH-B1 EPH-B4 EPH-A8 EPH-A2EPH-B2 EPH-B2 EPH-A2 EPH-B2 EPH-B2 FMS EPH-B1 EPH-A3 FMS EPH-B3 EPH-A3EPH-B4 EPH-B4 HIPK4 EPH-B2 EPH-A4 KIT EPH-B4 EPH-A4 ITK FMS KIT EPH-B3EPH-A5 LYNA FMS EPH-A5 JNK1 KIT KIT-D816V FMS EPH-A8 LYNB KIT EPH-A8JNK2 LYNA LCK KIT EPH-B1 PDGFR-α P38-α EPH-B1 JNK3 LYNB LYNA LYNA EPH-B2PDGFR-β P38-β EPH-B2 KIT P38-α MNK2 LYNB EPH-B3 ROS PDGFR-α EPH-B3 LYNAP38-β P38-α P38-α EPH-B4 TNK1 PDGFR-α EPH-B4 LYNB PDGFR-α P38-β P38-βFER FER P38-α PDGFR-β PDGFR-α PDGFR-α FES FES P38-β ROS PDGFR-β PDGFR-βFMS FMS PDGFR-α TNIK PI3K-α PI3K-α HCK HCK PDGFR-α TNK1 TNIK PLK1 HIPK4HIPK4 PI3K-α TNK1 ROS JNK1 JNK1 TEC TNIK JNK2 JNK2 TNK1 TNK1 JNK3 JNK3TXK KIT KIT LCK LCK LTK LTK LYNA LYNA LYNB LYNB MUSK MUSK P38-α P38-αP38-β P38-β PDGFR-α PDGFR-α PDGFR-β PDGFR-β PYK2 PYK2 ROS ROS TAOK3TAOK3 TIE2 TIE2 TNIK TNIK TNK1 TNK1

TABLE 27 Kinase Inhibition Profiles of the 20 new compounds defined inTable 25, continued. Inhibitors were profiled against a panel of 230kinaes when assayed at 5 uM. Scafffold # 15 16 17 25 26 27 29 50 51 5260 HGM# 35 35 35 35 35 35 35 35 35 35 35 Cpd # I-26 I-27 I-15 I-28 I-29I-30 I-31 I-32 I-33 I-34 I-35 AXL AXL BLK ABL1 AXL AXL BRK AXL AXL AXLCRAF BRK BLK BRK ARG BRK BLK CRAF BLK BLK DDR2 DDR2 CRAF BRK DDR2 AXLCRAF BRK DDR2 BRK DDR2 EPH-A4 EPH-A1 DDR2 CRAF EPH-A4 BLK DDR2 DDR2HIPK4 DDR2 EPH-A2 EPH-A5 EPH-A4 EPH-A4 DDR2 EPH-B2 BRK EPH-A1 EPH-A4 MEREPH-A2 EPH-A3 EPH-B4 EPH-A5 EPH-A5 EPH-A3 FMS CRAF EPH-A2 EPH-A5 MKNK1EPH-A3 EPH-A4 FLT3 EPH-A8 EPH-A8 EPH-A4 HIPK4 DDR2 EPH-A3 EPH-A8 MNK2EPH-A4 EPH-A5 HIPK4 EPH-B2 EPH-B1 EPH-A5 MER EPH-A1 EPH-A4 EPH-B2 MUSKEPH-A5 EPH-A8 KDR FGFR2 EPH-B2 EPH-A8 MNK2 EPH-A2 EPH-A5 EPH-B4 TIE2EPH-A8 EPH-B1 KIT FLT3 EPH-B4 EPH-B1 MUSK EPH-A3 EPH-A8 FLT3 TRKA EPH-B2EPH-B2 LCK FMS FER EPH-B2 SRMS EPH-A4 EPH-B2 FMS TRKC EPH-B4 EPH-B4 LOKHIPK4 FGR EPH-B4 TIE2 EPH-A5 EPH-B4 HCK TYRO3 FLT3 FLT3 MNK2 KDR FLT3FER TRKA EPH-A8 FGFR1 HIPK3 FMS HIPK4 MUSK KIT FMS FLT3 TRKB EPH-B2FGFR2 HIPK4 HIPK3 JAK2 P38-α P38-α HIPK4 FMS TRKC EPH-B4 FGFR4 JAK2HIPK4 KDR P38-β P38-β JAK2 HIPK4 TYRO3 FGR FLT1 KDR KDR KIT RET RET KDRJAK2 FLT1 FLT3 LCK KIT LCK SRC TAOK3 KIT KDR FLT3 FLT4 LOK LCK LOK TAOK2TIE2 LCK KIT FLT4 FMS LYNA LOK LYNA TAOK3 LOK LCK FMS HIPK4 LYNB MAP4K5LYNB TIE2 LYNA LOK HCK JAK2 MAP4K5 MER MAP4K5 TRKA MAP4K5 MAP4K5 HIPK3KDR MER MNK2 MER TYRO3 MEK2 MER HIPK4 KIT MET MUSK MNK2 YES MER MKNK1JAK2 LCK MNK2 P38-α MUSK MKNK1 MNK2 KDR LYNA MUSK P38-β P38-α MNK2 MUSKKIT LYNB PDGFR-α PDGFR-α P38-β MUSK P38-α LCK MAP4K5 PI3K-α SRC PI3K-αPDGFR-α PDGFR-α LOK MER RON TAOK3 RET RON TIE2 LYNA MKNK1 TIE2 TIE2TAOK2 TIE2 TRKA LYNB MNK2 TRKA TRKA TAOK3 TRKA TRKB MAP4K5 MUSK TRKBTRKB TIE2 TRKB TRKC MER P38-α TRKC TRKC TRKA TRKC TYRO3 MET PDGFR-αTYRO3 TYRO3 TRKB TYRO3 MKNK1 PDGFR-β YES TRKC MNK2 RET TYRO3 MUSK RONP38-α TIE2 PDGFR-α TRKA PDGFR-β TRKB PTK5 TRKC RON TYRO3 SRC TIE2 TRKATRKB TRKC TYRO3 YES

TABLE 28 Kinase Inhibition Profiles of Inhibitors I-36, I-37, and I-38against a panel of 250 kinaes when assayed at 5 uM. Scaffold # N-MethylN-Methyl S-17 S-17 S-17 HGM # HGM-11 HGM-35 HGM-1 Cpd ID# I-36 I-37 I-38ABL ARG ARG ARG AXL CRAF BRAF BLK DDR2 CLK4 BRAF FMS CRAF CRAF KIT CSKDDR2 MRCK-α DDR2 EPH-A3 PDGFR-α EPH-A2 EPH-A4 PDGFR-β EPH-A3 EPH-A5 ROSEPH-A4 EPH-A8 TNK1 EPH-A5 EPH-B2 EPH-A8 EPH-B4 EPH-B2 FER FLT-1 FGRFLT-2 FLT-2 FLT-3 FMS FMS HIPK4 KDR JAK2 KIT KDR LOK KIT LYNA LCK LYNBLOK MAP4K4 LYNA p38-α LYNB p38-β MKNK1 PDGFR-α MNK2 PDGFR-β MUSK PI3K-αPDGFR-α RET RON PIPK2 TIE2 TAOK2 TRKB TAOK3 TRKC TIE2 TYRO3

TABLE 29 Kinase Inhibition Profiles of Inhibitors I-41 to I-51 against apanel of 250 kinaes when assayed at 5 uM. Scaffold # S-84 S-84 S-84 S-84S-52 S-84 Sulfoxide Sulfone S-81 S-82 S-83 S-17 S-84 Sulfoxide SulfoneHGM # HGM-7 HGM-7 HGM-7 HGM-7 HGM-7 HGM-7 HGM-7 HMG-4 HMG-4 HMG-4 HMG-4Cpd ID# I-41 I-42 I-43 I-44 I-45 I-46 I-47 I-48 I-49 I-50 I-51 BRAF ARGBRAF CRAF BRAF BRAF ABL ARG ABL1 ABL1 ARG CRAF BLK CRAF DDR2 CRAF CRAFARG CRAF ARG ARG BRAF DDR2 BMX DDR2 EPH-A8 DDR2 DDR2 BRAF DDR2 AURORA-AAURORA-A CRAF EPH-A2 BRAF EPH-A2 FMS EPH-A2 EPH-A2 CRAF EPH-A2 BLK BLKDDR2 EPH-A3 BRK EPH-A3 p38-α EPH-A4 EPH-A4 CSK EPH-A3 BMX BMX EPH-A2EPH-A4 CRAF EPH-A4 PDGFR-α EPH-A5 EPH-A5 DDR2 EPH-A4 BRAF BRAF EPH-A3EPH-A5 CSK EPH-A5 PDGFR-β EPH-A8 EPH-A8 EPH-A2 EPH-A5 BRK CRAF EPH-A4EPH-A8 DDR2 EPH-A8 EPH-B2 EPH-B2 EPH-A4 EPH-A8 BTK DDR2 EPH-A5 EPH-B1EPH-A1 EPH-B2 FLT-1 KIT EPH-A5 EPH-B2 CAMK1A EPH-A2 EPH-A8 EPH-B2 EPH-A2EPH-B4 FLT-3 LCK EPH-A8 KDR CLK4 EPH-A3 EPH-B2 EPH-B4 EPH-A3 FLT-1 FMSLYNA EPH-B2 KIT CRAF EPH-A4 FER FLT-1 EPH-A4 FLT-3 KDR LYNB FMS LYNB CSKEPH-A5 FLT-1 FLT-3 EPH-A5 FMS KIT p38-α FYN p38-α DDR2 EPH-A8 FLT-3 FMSEPH-A8 KDR LYNA p38-β HCK PDGFR-α EPH-A1 EPH-B1 FMS KDR EPH-B1 KIT LYNBPDGFR-α KDR PDGFR-β EPH-A2 EPH-B2 KDR KIT EPH-B2 LYNB p38-α PDGFR-β KITPI3K-α EPH-A3 EPH-B3 KIT LYNA EPH-B3 p38-α p38-β RET LCK PI3K-γ EPH-A4EPH-B4 LYNA LYNB EPH-B4 p38-β PDGFR-α LYNA EPH-A5 FER LYNB p38-α FGFR1PDGFR-α LYNB EPH-A8 FGFR4 MAP4K2 p38-β FGR PDGFR-β p38-α EPH-B1 FGRMAP4K5 PDGFR-α FLT-1 p38-β EPH-B2 FLT-1 NEK1 PDGFR-β FLT-3 PAK4 EPH-B3FLT-3 p38-α FMS PDGFR-α EPH-B4 FMS p38-β FYN PDGFR-β ERB-B4 FYN PDGFR-αHCK RET FAX HCK PDGFR-β KDR PIPK2 FER KDR PI3K-α KIT SRC FGFR1 KIT TAOK3LCK YES FGFR2 LCK TIE2 LYNA FGFR3 LOK LYNB FGFR4 LYNA MAP4K2 FGR LYNBMAP4K5 FLT-1 MAP4K2 NEK1 FLT-2 MAP4K5 p38-α FLT-3 MUSK P38-β FMS NEK1PDGFR-α FYN p38-α PDGFR-β HCK P38-β RET HIPK4 PDGFR-α PIPK2 KDR PDGFR-βSRC KIT RET TAOK2 LCK TAOK3 TAOK3 LOK TIE2 TIE2 LYNA TNK1 LYNB YESMAP4K2 MAP4K4 MAP4K5 MELK MINK MUSK NEK1 p38-α p38-β PDGFR-α PDGFR-βPIM3 PTK5 PYK2 RET PIPK2 ROS SRC SRMS TAK1-TAB1 TAOK2 TAOK3 TIE2 TNIKTNK1 TRKB TRKC TXK YESAdditional Uses for Kinase Inhibitors with this Scaffold Geometry:New Tools to Identify Kinases Capable of Atypical Conformations:

Taken together, these data demonstrate the broad applicability andutility of this scaffold geometry for the targeted inhibition of a largenumber of kinases. Therefore, because this scaffold geometry is believedto be highly complementary to the conserved topography of lowerselectivity sites created by movements of a kinase's DFG loop, compoundsbased on this generic geometry may be expected to induce or stabilizethese conformational changes particularly well. Indeed, kinases thathave been previously not been known to adopt these conformations mayprove amenable to inhibition by or association with inhibitors built onthis scaffold concept. For example, affinity reagents that target aspecific inactive form of protein kinases have been described and usedto access if a kinase can adopt this inactive conformation, (Ranjitkar,2010).

Similarly, inhibitors and affinity agents derived from molecules thatshare the specific scaffold geometry described herein; such asInhibitors I-12, I-14, I-15, and the like; will have applications asnovel tools for the identification of kinases that can adopt inactive orother atypical conformations. Once selected, the specific structuralinformation could then be confirmed by X-ray co-crystal structuredetermination. Examples of such inhibitors and their correspondingaffinity probes are illustrated in FIG. 15, which shows representativeinhibitors and analogous affinity probes useful for the identificationof atypical kinase conformations where R₀ is as defined above.

The interesting profiles and a wide range of selectivity demonstrated bythese few early compounds support many utilities for kinase inhibitorsthat share this scaffold geometry. These data suggest that incorporationof Hinge-Gatekeeper Motifs from either known type-II inhibitors or fromATP competitive inhibitors are expected to provide novel kinaseinhibition profiles that may have therapeutic potential. Anotherpotential utility for compounds sharing this scaffold geometry is forthe evaluation and identification of the potential for a given kinase toassume an atypical or DFG-out conformation. A third utility for kinaseinhibitors that share this scaffold geometry is their adaptation forincorporation of radio labels for positron emission tomography,

Adaptation of this Scaffold to the Preparation Kinase-Targeted PETImaging Agents:

In contrast to recently reported cumbersome methods described for theintroduction of ¹⁸F into kinase inhibitors (Griffiths, 2010), we hereindescribe several novel, generally applicable, and highly practicalsynthetic strategies for the incorporation of ¹⁸F or ¹¹C labels in thelast step(s) of PET probe preparation FIG. 16.

Two precedents are illustrated for the introduction of fluoride in FIG.16. In the first, a cyclic sulfamate 1 is used as the activatedintermediate for fluoride displacement. In the second an imidazoylsulfonate 2 provides the necessary activation. Application of the secondprecedent to an appropriate 2-hydroxymethyl scaffold should provide theradiolabeled derivative 5 via imidazoyl-sulfonate 3, procedure-A.Procedure-B is a special case for selected imidazole scaffolds in whichaddition of imidazole is not required as formation of an intramolecularcyclic imidazoly sulfonate 4 should be formed. Intermediate 4 could beformed and used immediately for introduction of [18]F label or storedfor later use. Due to the 110 minute half-life of [18]F(http://www.iem-inc.com/toolhalf.html), the strength of this strategy isthat it occurs at the end of the synthesis and allows production of the[18]F probes 5 and 6 in a single step just prior to utilization.

Due to the even shorter 20 minute half life of [11]C, utilization ofthis radio-isotope is more challenging. Nevertheless, our observationthat inhibitors that incorporate the 2-methylsulfonyl 5-memberedheteroaryl carboxamide scaffold display attractive selectivity profilesprovides a viable alternative for the rapid introduction of [11]Cradio-label. Using a fully elaborated 2-mercapto-thiazole precursor 7,alkylation with [11]C methyl iodide or bromide would quickly provide the2-methyl mercapto inidazole 8 in good yield (Garcia, 2005; Tang, 2011).If required, the corresponding sulfoxide or sulfone analogs could beoptionally prepared or, depending on metabolism, in vivo oxidation mayprovide these oxidation states Due to the short half lives of both [18]Fand [11]C, introduction of the radiolabel as the end of the synthesisoffers a clear advantage.

Alternate strategies for the introduction of [18]F into an ary ring suchas PhenoFluor™(1,3-Bis(2,6-diisopropylphenyl)-2,2-difluoro-2,3-dihydro-1H-imidazole)is commercially available (Sigma-Aldrich product directory as catalognumber SFL0001) and could be exploited using an appropriate phenolicintermediate (see http://www.scifluor.com), FIG. 17.

Uniqueness as a Platform Technology:

Based on our recent findings, Tables 3 and 4, the 2-substitutedortho-aryl-imidazole carboxamide scaffold is broadly recognized, albeitmodestly in some cases, by a wide variety of kinases. Althoughco-crystal structure determinations will be needed to confirm the modeof binding for each kinases, many of the inhibited kinases are thoughtto be capable of adopting the DGF-out conformation. In addition to beingwidely accepted, this scaffold is proposed to adopt a novel mode ofbinding within the conserved topography of the lower selectivity-siteproduced by movement of the DFG loop from its pocket, FIG. 10. In sodoing, a highly complementary fit provides a uniquely rigid anchor pointagainst which distances between affinity and selectivity elements aredefined. The selectivity for p38-α observed for Inhibitor 11, Table-3,may be an example of this rigid anchoring function.

Taken together, this invention describes a new platform technology thatcan be useful for both the discovery of new targeted therapeutics aswell as the development of highly selective PET imaging agents. Thecapability to achieve both broad applicability and to dial inexceptional selectivity with a single scaffold is highly unusual.Moreover, because the scaffolds and the chemistries described hereincan, in principle, also be adapted to the preparation of structurallysimilar PET imaging probes, this invention offers unique opportunitiesfor synergy and cost savings, FIG. 18. For example, the ability todevelop selective imaging agents for kinase targets will be particularlyuseful to establish proof of concept from preclinical animal studiesthrough clinical evaluation. In addition, selection of the mostappropriate patient populations for efficacy studies will likely resultin significant cost savings (de Vries, 2010). Eventually, application ofimaging agents based on this generic technology will be used for thediagnosis, characterization, and monitoring of diseases characterized bythe over expression, up-regulation, mutation, and aberrant activity ofkinases.

The 2-substituted 5-aryl-imidazole carboxamide scaffold has been shownto be both unique and broadly applicable to the design of multi-targetedtype-II kinase inhibitors. The unique geometry defined by this scaffoldis believed to be particularly well suited to establish novelinteractions at the bottom of the lower selectivity site that can beexploited to increase binding affinity, rigidity, and selectivity. FIG.18 illustrates various strategies for construction of penultimateintermediates or final kinase inhibitors and imaging agents using thescaffold platform described herein. Scaffold acids can be coupled to asuitably protected HGM amine (Route-1); to a suitably protectedgatekeeper residue followed by deprotection and coupling of andappropriately protected hinge-linger group (Route-2); or to a suitablyprotected linker-gatekeeper fragment followed by deprotection andcoupling to a suitably protected hinge interacting group.

It is proposed that the aryl ring of the 5-membered heteroarylcarboxamide scaffold is ideally positioned to closely mimic interactionsnormally made by the phenylalanine side-chain of the DFG-loop.Therefore, when an appropriate Hinge-Gatekeeper interacting substitutent(H)-L-G- is attached to an appropriate ortho-aryl 5-membered heteroarylcarboxamide scaffold as in Formula-1, the unique geometry defined bythis scaffold results in a highly complementary and tight fit with thenew lipophilic pocket (lower selectivity site) generated as a result ofthe DFG-loop movement. This invention describes a previouslyunappreciated common feature of the bottom of the lower selectivity sitethat can be exploited by an appropriate substituent on the Ar group inFormula-I to provide both affinity and selectivity enhancements. Thisconserved feature or indentation results from the common method ofconstruction of this lipophilic pocket in many kinases.

Because we have demonstrated that a scaffold with this combination offunctionality and scaffold geometry can both yield broad applicabilityand yet, with simple modification, lead to a highly selective inhibitor,this scaffold geometry is considered a general platform technology forthe stabilization or inducement of the inactive DFG-out conformation ofkinases capable of adopting it. Molecular modeling studies suggest thatthe binding mode described herein is consistent with both the observedSAR and the observed broad applicability conferred by an imidazolescaffold that shares this general geometry.

Our observation that the indentation formed by convergence of aliphaticamino acid side chains is a general feature at the bottom of the lowerselectivity site of many kinases is new. However, interactions describedin these models need not be the only mode of binding by which kinaseinhibition can result nor is the inactive DFG-out conformation,necessarily, the only kinase conformation that these inhibitors may bindto.

Compounds that target a combination of tyrosine kinases that areassociated with oncogene addiction that also block escape orcompensatory mechanisms shown to lead to tyrosine kinase inhibitor (TKI)drug resistance would be of high value. We have demonstrated that accessto unique selectivity profiles can be achieved using the scaffoldconcepts described herein. For example, compounds SFE-00013 and SFE-0014inhibit PI3Ks in addition to Raf (including B-Raf, B-Raf mutant V600E),C-Raf, p38-α and PDGFR-α, thereby simultaneously inhibiting both thecritical Ras/Raf/MEK/ERK and PI3K/AKT/mTOR pathways. On the other hand,SFE-0011 is very selective for p38-α but with modest PI3K activity.Optimization of this lead may lead to a novel approach for the treatmentof chronic inflammation. It is expected that these and other uniqueprofiles resulting from this invention will provide both noveltherapeutic agents for the treatment of cancers, pain, infectious andneurodegenerative diseases, and inflammation related conditions.

The variety of kinases inhibited and the variations in the inhibitionprofiles produced by both variation of the HGM and the scaffoldfunctionality illustrates the utility of this new scaffold geometry forthe identification of novel agents useful for the treatment of diseaseswhere the mutation, over expression, or aberrant activity of kinasesthrough loss of phosphatase activities play a contributory role.

In general, PET imaging agents with targeted kinase inhibitor activitywill be allow the study of roles kinase targets play in disease relatedphysiology. In a special case, compounds the represent the subset ofFormula I where R₀ is defined as —CH₂[18]F or —SO_(m)[11]CH₃ provideunique utilities as positron emission tomography (PET) imaging agentsfor the study, diagnosis, characterization, and monitoring of diseasesand conditions resulting from the aberrant expression, activation, oractivity of the kinases for which they inhibit. Moreover, thetechnologies for the general introduction of radio label describedherein, constitute a generic technology for the preparation and routineproduction of selective multi-target kinase inhibitor-based imagingagents.

It is anticipated that the combination of appropriately targeted PETimaging agents combined with the availability of either closely relatedinhibitors or structurally different inhibitors with equivalentselectivity profiles offers an innovative and useful strategy for theidentification and development of patient specific therapies forconditions where the aberrant expression, activation, or mutation of akinase or profile of kinases is indicated.

What is claimed is:
 1. A method of treating cancer, restenosis afterpercutaneous coronary intervention, venous bypass graft disease, type-2diabetes, or neuropathic pain, comprising administering to a humansubject an effective amount of a compound represented by structuralFormulae (VII) to (XIV):

or a pharmaceutically acceptable salt, wherein: 1) L is —O—, —S—, —NH—or —C(O)NR¹; and (H) is of the structural formulae

or 2) L is —O—, —NH—, —CC—, or —C(O)NR¹; and (H) is of the structuralformulae

R₁₁ is selected from H, —CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃,and —S—CH₃; R₀ is selected from —F, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹, —OR²,—CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F,—CF[18]F₂, and —C[18]F₃; provided that R⁰ is not CF₃ when the ring towhich it is attached is a 1H-imidazole ring; R¹ is independently H, CH₃,—CH₂CH₃, or cyclopropyl; R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or—(CH₂)_(n)—NR^(A)R^(B); R^(A) and R^(B) are each independently H, CH₃,—CH₂CH₃, or cyclopropyl; or R^(A) and R^(B) taken together form a 3-6membered carbocyclic ring system or 5-7 membered saturated heterocyclicring system; Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹,—NR^(A)R^(B), —NR¹COR², —S(O)_(m)R¹, —SO₂NR^(A)R^(B),—[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃, —[O—(CH₂)₂]_(n)—CH₂ ¹⁸F, —CH₂—F;—CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F, —CF[18]F₂, or—C[18]F₃; m is 0, 1, or 2; n is 1, 2, or 3; U is —H, F, Cl, —OR¹, or—NHR¹; V and W are each independently selected from —H, —F, —Cl, —CF₃,—CONHR², —X—R¹, —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹, X—(CH₂)_(m)CONR¹R²,—X—CH₂—(CH₂)_(n)OR, —X—CH₂—(CH₂)_(n)NR¹R², —X—CH₂—(CH₂)_(n)S(O)_(m)R¹,—X—(CH₂)_(n)S(O)_(m)NR¹R², —O—(CH₂)₂NR¹R², —O—(CH₂)₃NR¹R²,—O—(CH₂)_(n)CONR¹R², —C₅-C₆heteroaryl, —COCH═CH—(CH₂)_(n)NR¹R², and T; Tis:

Z is independently selected from —CH—, —CF—, and —N—; R₄ is selectedfrom H, —CH₃, —CH₂CH₃, cyclopropyl, —F, -[18]F, —Cl, —Br, —CH₂F,—CH₂[18]F, —CHF₂, —CF₃, —HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and—S(O)_(m)R¹; and X is —O—, —NR²—, or —S—.
 2. A method of treatingcancer, restenosis after percutaneous coronary intervention, venousbypass graft disease, type-2 diabetes, or neuropathic pain, comprisingadministering to a human subject an effective amount of a compoundrepresented by structural Formulae (XX) to (XXVIV):

or a pharmaceutically acceptable salt thereof, wherein: 1) L is —O—,—S—, —NH— or —C(O)NR¹; and (H) is of the structural formulae

or 2) L is —O—, —NH—, —CC—, or —C(O)NR¹; and (H) is of the structuralformulae

R₁₁ is selected from H, —CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃,and —S—CH₃; R₀ is selected from —F, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹, —OR²,—CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F,—CF[18]F₂, and —C[18]F₃; R¹ is independently H, CH₃, —CH₂CH₃, orcyclopropyl; R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or—(CH₂)_(n)—NR^(A)R^(B); R^(A) and R^(B) are each independently H, CH₃,—CH₂CH₃, or cyclopropyl; or R^(A) and R^(B) taken together form a 3-6membered carbocyclic ring system or 5-7 membered saturated heterocyclicring system; Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹,—NR^(A)R^(B), —NR¹COR², —S(O)_(m)R¹, —SO₂NR^(A)R^(B),—[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃, —[O—(CH₂)₂]_(n)—CH₂ ¹⁸F, —CH₂—F;—CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F, —CF[18]F₂, or—C[18]F₃; m is 0, 1, or 2; n is 1, 2, or 3; U is —H, F, Cl, —OR¹, or—NHR¹; V and W are each independently selected from —H, —F, —Cl, —CF₃,—CONHR², —X—R¹, —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹, X—(CH₂)_(m)CONR¹R²,—X—CH₂—(CH₂)_(n)OR, —X—CH₂—(CH₂)₂NR¹R², —X—CH₂—(CH₂)_(n)S(O)_(m)R¹,—X—(CH₂)_(n)S(O)_(m)NR¹R², —O—(CH₂)₂NR¹R², —O—(CH₂)₃NR¹R²,—O—(CH₂)_(n)CONR¹R², —C₅-C₆heteroaryl, —COCH═CH—(CH₂)_(n)NR¹R², and T; Tis:

Z is independently selected from —CH—, —CF—, and —N—; R₄ is selectedfrom H, —CH₃, —CH₂CH₃, cyclopropyl, —F; -[18]F, —Cl, —Br, —CH₂F,—CH₂[18]F, —CHF₂, —CF₃, —HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and—S(O)_(m)R¹; and X is —O—, —NR²—, or —S—.
 3. A method of treatingcancer, restenosis after percutaneous coronary intervention, venousbypass graft disease, type-2 diabetes, or neuropathic pain, comprisingadministering to a human subject an effective amount of a compoundrepresented by structural Formulae (VII) to (XIV):

or a pharmaceutically acceptable salt, wherein: L is of the structuralformulae

(H) is of the structural formulae

R₁₁ is selected from H, —CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃,and —S—CH₃; R₀ is selected from —F, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹, —OR²,—CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F,—CF[18]F₂, and —C[18]F₃; provided that R⁰ is not CF₃ when the ring towhich it is attached is a 1H-imidazole ring; R¹ is independently H, CH₃,—CH₂CH₃, or cyclopropyl; R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or—(CH₂)_(n)—NR^(A)R^(B); R^(A) and R^(B) are each independently H, CH₃,—CH₂CH₃, or cyclopropyl; or R^(A) and R^(B) taken together form a 3-6membered carbocyclic ring system or 5-7 membered saturated heterocyclicring system; Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹,—NR^(A)R^(B), —NR¹COR², —S(O)_(m)R¹, —SO₂NR^(A)R^(B),—[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃, —[O—(CH₂)₂]_(n)—CH₂18F, —CH₂—F;—CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F, —CF[18]F₂, or—C[18]F₃; m is 0, 1, or 2; n is 1, 2, or 3; V is selected from —H, —F,—Cl, —CF₃, —CONHR², —X—R¹, —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹,X—(CH₂)_(m)CONR¹R², —X—CH₂—(CH₂)_(n)OR¹, —X—CH₂—(CH₂)_(n)NR¹R²,—X—CH₂—(CH₂)_(n)S(O)_(m)R¹, —X—(CH₂)_(n)S(O)_(m)NR¹R², —O—(CH₂)₂NR¹R²,—O—(CH₂)₃NR¹R², —O—(CH₂)_(n)CONR¹R², —C₅- C₆heteroaryl,—COCH═CH—(CH₂)_(n)NR¹R², and T; T is:

Z is independently selected from —CH—, —CF—, and —N—; R₄ is selectedfrom H, —CH₃, —CH₂CH₃, cyclopropyl, —F; -[18]F, —Cl, —Br, —CH₂F,—CH₂[18]F, —CHF₂, —CF₃, —HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and—S(O)_(m)R¹; R¹⁰ is hydrogen; and X is —O—, —NR²—, or —S—.
 4. A methodof treating cancer, restenosis after percutaneous coronary intervention,venous bypass graft disease, type-2 diabetes, or neuropathic pain,comprising administering to a human subject an effective amount of acompound represented by structural Formulae (XX) to (XXVIV):

or a pharmaceutically acceptable salt thereof, wherein: L is of thestructural formulae

(H) is of the structural formulae

R₁₁ is selected from H, —CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃,and —S—CH₃; R₀ is selected from —F, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹, —OR²,—CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F,—CF[18]F₂, and —C[18]F₃; R¹ is independently H, CH₃, —CH₂CH₃, orcyclopropyl; R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or—(CH₂)_(n)—NR^(A)R^(B); R^(A) and R^(B) are each independently H, CH₃,—CH₂CH₃, or cyclopropyl; or R^(A) and R^(B) taken together form a 3-6membered carbocyclic ring system or 5-7 membered saturated heterocyclicring system; Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹,—NR^(A)R^(B), —NR¹COR², —S(O)_(m)R¹, SO₂NR^(A)R^(B),—[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃, —[O—(CH₂)₂]_(n)—CH₂18F, —CH₂—F;—CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F, —CF[18]F₂, or—C[18]F₃; m is 0, 1, or 2; n is 1, 2, or 3; V is selected from —H, —F,—Cl, —CF₃, —CONHR², —X—R¹, —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹,X—(CH₂)_(m)CONR¹R², —X—CH₂—(CH₂)_(n)OR¹, —X—CH₂—(CH₂)_(n)NR¹R²,—X—CH₂—(CH₂)_(n)S(O)_(m)R¹, —X—(CH₂)_(n)S(O)_(m)NR¹R², —O—(CH₂)₂NR₁R²,—O—(CH₂)₃NR¹R², —O—(CH₂)_(n)CONR¹R², —C₅- C₆heteroaryl,—COCH═CH—(CH₂)_(n)NR¹R², and T; T is:

Z is independently selected from —CH—, —CF—, and —N—; R₄ is selectedfrom H, —CH₃, —CH₂CH₃, cyclopropyl, —F; -[18]F, —Cl, —Br, —CH₂F,—CH₂[18]F, —CHF₂, —CF₃, —HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and—S(O)_(m)R¹; R¹⁰ is hydrogen; and X is —O—, —NR²—, or —S—.
 5. A methodof treating cancer, restenosis after percutaneous coronary intervention,venous bypass graft disease, type-2 diabetes, or neuropathic pain,comprising administering to a human subject an effective amount of acompound selected from the formulae:

or a pharmaceutically acceptable salt thereof.
 6. A method of treatingcancer, restenosis after percutaneous coronary intervention, venousbypass graft disease, type-2 diabetes, or neuropathic pain, comprisingadministering to a human subject an effective amount of a compoundselected from the formulae:

or a pharmaceutically acceptable salt thereof.
 7. A method of treatingcancer, restenosis after percutaneous coronary intervention, venousbypass graft disease, type-2 diabetes, or neuropathic pain, comprisingadministering to a human subject an effective amount of a compoundrepresented by structural Formulae (VII) to (XIV):

or a pharmaceutically acceptable salt, wherein: 1) L is a bond, —O—,═NH—, —S(O)_(m)—, —CC—, —O—CHR¹—, —NHCHR¹, S(O)_(m)CHR¹, —CHR¹O—,—CHR¹NH—, or —CHR¹S(O)_(m)—; and (H) is of the structural formulae

R₁₁ is selected from H, —CH₃, —CH₂CH₃, cyclopropyl, —F, —Cl, —CN, —OCH₃,and —S—CH₃; R₀ is selected from —F, —C₂-C₄ alkenyl, —C₂-C₄ alkynyl,—CO(CH₂)_(n)Y, —(CH₂)_(n)—NR^(A)R^(B), —[O—(CH₂)₂]_(n)Y,—(CH₂)_(n)SO₂NR^(A)R^(B), —S(O)_(m)—(CH₂)_(m)—R¹, —S(O)_(m)R¹, —OR²,—CH₂—F, —CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F,—CF[18]F₂, and —C[18]F₃; provided that R⁰ is not CF₃ when the ring towhich it is attached is a 1H-imidazole ring; R¹ is independently H, CH₃,—CH₂CH₃, or cyclopropyl; R² is independently H, —CH₃, —(CH₂)_(n)—CH₃, or—(CH₂)_(n)—NR^(A)R^(B); R^(A) and R^(B) are each independently H, CH₃,—CH₂CH₃, or cyclopropyl; or R^(A) and R^(B) taken together form a 3-6membered carbocyclic ring system or 5-7 membered saturated heterocyclicring system; Y is —CHR¹R², —CN, —COR¹, —CONR^(A)R^(B), —OR¹,—NR^(A)R^(B), —NR¹COR², —S(O)_(m)R¹, —SO₂NR^(A)R^(B),—[O—(CH₂)₂]_(n)—CH₂F, —S(O)_(m)[11]CH₃, —[O—(CH₂)₂]_(n)—CH₂ ¹F, —CH₂—F;—CH₂[18]F, —CHF₂, —CHF[18]F, —CH[18]F₂, —CF₃, —CF₂[18]F, —CF[18]F₂, or—C[18]F₃; m is 0, 1, or 2; n is 1, 2, or 3; U is —H, F, C1, —OR¹, or—NHR¹; V and W are each independently selected from —H, —F, —Cl, —CF₃,—CONHR², —X—R¹, —X—(CH₂)_(n)CN, —X—(CH₂)_(m)COR¹, X—(CH₂)_(m)CONR¹R²,—X—CH₂—(CH₂)_(n)OR, —X—CH₂—(CH₂)_(n)NR¹R², —X—CH₂—(CH₂)_(n)S(O)_(m)R¹,—X—(CH₂)_(n)S(O)_(m)NR¹R², —O—(CH₂)₂NR¹R², —O—(CH₂)₃NR¹R²,—O—(CH₂)_(n)CONR¹R², —C₅-C₆heteroaryl, —COCH═CH—(CH₂)_(n)NR¹R², and T; Tis:

Z is independently selected from —CH—, —CF—, and —N—; R₄ is selectedfrom H, —CH₃, —CH₂CH₃, cyclopropyl, —F, -[18]F, —Cl, —Br, —CH₂F,—CH₂[18]F, —CHF₂, —CF₃, —HC═CHR¹, —CCR¹; —CN, —OCF₃, —NHR¹—OR¹, and—S(O)_(m)R¹; and X is —O—, —NR²—, or —S—.